N-myristoylation: An Overview

N-myristoylation: An Overview N-myristoylation Protein N-myristoylation is the covalent attachment of myristate, a 14-carbon fatty acid, onto the N-terminal glycine residues of protein substrates. It is transferred co- or post-translationally to a subset of proteins from a thioester form, myristoyl-CoA, catalyzed by N-myristoyl transferases (NMTs). (insert genes expressing nmt1 and nmt2, NMT recognizes a general consensus sequence for myristoylation (Gly-X-X-X-(Ser/Thr/Cys)) containing a N-terminal glycine, 3 other amino acids and a serine, threonine or a cysteine in the fifth position.) While this process is often observed co-translationally on nascent shorter protein substrates, post-translation myristoylation ensues during apoptosis on N-terminal glycine residue exposed after caspase cleavage of protein substrates.1 An increase in proteins hydrophobicity conferred by this modification allows for weak protein-lipid and protein-protein interactions, as well as for membrane targeting and function of proteins involved in signal tr ansduction cascades.2 NMTs served as therapeutic targets owing to their importance for the survival of human pathogens and their association with carcinogenesis.1 To globally profile NMT protein substrates, chemical proteomic approaches have been employed where small tags on fatty acids such as an alkyne (YnMyr) or azide (AzMyr) (Fig. X) were developed to probe myristoylated proteins via metabolic labeling.3,4 This probe was successfully used in a high-confidence profiling of the co-translational myristoylome in human and zebra fish.5 Although YnMyr remains to be the probe of choice owing to its minimal background labeling6, it was demonstrated to label proteins with other known lipid-modifications such as NÃŽÂ µ-myristoyl,7 S-palmitoyl8 and GPI-anchors9-compromising its specificity towards labeling of N-terminal myristoylated proteins. To circumvent the challenge of identifying the true NMT substrates, Tate et al. used an integrated chemical biology approach where selective inhibition of NMT with smal l-molecule inhibitors combined with YnMyr labeling and quantitative proteomics allowed for profiling of more than 30 known and novel protein candidates for N-myristoylation in blood-stage malaria parasite.9 (describe that the presence of inhibitor abolished the labeling of the true substrates, which should not be enriched in control samples) This technology was also applied to globally profile the N-myristoylome of other human pathogens such as in  Leishmania donovani,10 Trypanosoma brucei,11and recently Trypanosoma cruzi.12Theprofiling of a large set of N-myristoylated proteins with diverse cellular functions unravels the significance of this lipid modification in these parasites. Furthermore, this also validates NMT as a viable drug target in attenuating the virulence of these pathogens. Extending the same approach to HeLa cancer cells enabled the identification of more than 100 of both co- and post-translationally modified N-myristoylated proteins, majority of which were identi fied at endogenous levels for the first time.13 Indeed, this robust technique proved to be powerful in discriminating on-target proteins from off-targets in a proteome-wide analysis, resulting in the discovery of novel NMT protein substrates at high confidence. Although promising, the method described where NMT inhibitors were used may not be applicable to more complex systems where cell viability may be compromised, e.g. in the context of viral and bacterial infection. An alternative targeted approach tosimplify data analysis of enriched proteins employs isolating those that bear the N-terminal glycine requirement for N-myristoylation. This enabled the profiling of downregulated host N-myristoylated proteins upon infection with herpes simplex virus (HSV)14, as well as novel fatty-acylated proteins encoded by HSV. This same approach provided a more defined picture of the demyristoylating function of the bacterial effector IpaJ upon host cell invasion of Shigella flexneri, which was determined to contribute to its virulence.15   Palmitoylation Proteins S-palmitoylation is the attachment of a 16-carbon long fatty acid (as palmitate-CoA) to cysteine residues, which was first discovered by radiolabeling of virus-infected cells with [3H]palmitate.16 The formation of the thioester linkage is mediated by a family of protein acyl transferases (PATs) that bear a conserved Asp-His -His-Cys catalytic motif (DHHC-PATs), which can be removed by hydrolysis aided by acyl protein thioesterases (APTs).17 Owing to the reversibility of this modification, S-palmitoylation of proteins was thought to be dynamically regulated ,whereby a subset of proteins are transiently palmitoylated in a certain time point/cellular activity. (insert something) S-palmitoylation has been demonstrated to be an essential mechanism for protein stability, activity, and proper cellular localization.18 Recent advances in identifying palmitoylated proteins revealed not only its key role in regulatory mechanisms but as well as in host invasion and virulence of pathogen s. Large-scale proteomic profiling of S-palmitoylated proteins using metabolic labeling has been heavily dependent on employing the alkyne analogue of palmitic acid, 17-ODYA (Fig. X). This commercially available chemical reporter is suitable for these analyses as it has shown better specificity and has minimal background in labeling proteins that are ought to be acylated by shorter fatty alkyl chains.6 The subsequent click reaction with fluorophore- or biotin-azide then allows for in-gel fluorescence monitoring and biotin-pulldown strategy prior to LC-MS proteomic analysis of labeled proteins, respectively. In these studies, hundreds of palmitoylated proteins were identified with a wide range of functions, highlighting the importance of S-palmitoylation in a plethora of cellular mechanisms and pathways. For instance, the first report on using such strategy applied to mammalian cells identified around 125 candidate S-palmitoylated proteins at high confidence, including G proteins, recept ors and uncharacterized hydrolases.19 Using the same strategy in dendritic immune cells (DC2.4) identified more than 150 predicted S-palmitoylated proteins and revealed that palmitoylation of interferon-induced transmembrane protein 3 (IFITM3)20 and Toll-like receptor 2 (TLR2)21 is essential for their antiviral activity. A more recent study on Cryptococcus neoformans revealed that a single PAT, Pfa4, palmitoylates the fungal proteins required for parasite integrity and virulence-palmitoylating 72 proteins identified in a global-scale approach.22 A more quantitative approach to measure levels of palmitoylated proteins combines metabolic labeling with 17-ODYA and Stable Isotope Labeling with amino acids in Cultured Cells (SILAC). In virus-infected RPE-1 epithelial cells, selective repression was observed for host S-palmitoylated proteins, including interferon signaling regulators and members of the tetraspanin family.14 A novel set of HSV-encoded proteins palmitoylated by the host machinery were selectively and significantly identified, further suggesting that HSV exploits the palmitoylation pathway which contributes to its virulence. As palmitoylation is a reversible process, the dynamic cycling of palmitoylated proteins in mouse T-cell hybridoma cells was investigated using this quantitative approach in combination with a pulse-chase technique.23 Through the use of a serine lipase-inhibitor as the chase, palmitoylated proteins that undergo fast turnovers were distinguished from those that are stably modified. This indicates that a subset of this dynamic palmitoylation event is regulated by serine hydrolases, validating the fundamental regulatory mechanism of depalmitoylation for proteins with rapid turnovers. It is important to note that in this study, only the insoluble protein fractions were analyzed, as the soluble proteins were not amenable to metabolic probe incorporation.19 Given the dynamic nature of palmitoylation, the metabolic labeling strategy would allow labeling of only those that are palmitoylated at the time of probe treatment and were stably modified. An older approach, coined as acyl-biotin exchange (ABE), has the potential to capture the full complement of palmitoylated proteins. In this multistep procedure, the protein lysates are treated with hydroxylamine to selectively cleave the thioester bonds, followed by disulfide capture with thiol-containing biotin analogue, and subsequently enriched through a pulldown technique prior to LC-MS analysis. ABE was first utilized in tandem with semi-quantitative MudPit analysis on profiling the palmitome of Saccharomyces cerivisae.24 The 12 known and 35 new palmitoylated proteins identified presented the first evidence on the diverse specificities of PATs. The ABE method was further employed in profiling the palmitoylome in rat neurons,25 human T cells,26 and recently in poplar tree cells,27 establishi ng its applicability to a wide range of biological systems. Both ABE and metabolic labeling approaches combined with SILAC revealed their large complementarity in profiling S-palmitoylated proteins in Plasmodium falciparum.28 A total of more than 400 palmitoylated proteins were identified where 202 proteins were enriched in both methods. As expected, metabolic labeling identified a lesser number of proteins, reflecting the less complexity in this approach. A pulse-chase labeling using ABE in a quantitative approach with 2-BMP as the parasite PAT inhibitor revealed the identification of a range of stably and dynamically palmitoylated proteins. Indeed, this study demonstrated the importance of palmitoylation in multiple parasite-specific processes, specifically in drug resistance, asexual stage development, host cell invasion, and protein export. Both methods were also employed in investigating the dysregulation of palmitoylation in breast cancer cells by inducing Snail-overexpression- an event correlated with chemoresistance and metastasis.29 Results showed that some proteins were differentially expressed regardless of differential palmitoylation. Thus, Snail-overexpression compromises the dynamic palmitoylation of some proteins that may be involved in pathways that contribute to malignancy. Albeit most proteins are S -palmitoylated in their cysteine residues, others were reported to be O-palmitoylated30 and N-palmitoylated17, which are also labeled by 17-ODYA. To distinguish S-palmitoylated proteins from these other forms in Toxoplasma gondii, a method similar to ABE was employed which also takes advantage of the labilityof thioester bonds to hydrolysis.31 In this approach, the metabolic incorporation of 17-ODYA and enrichment is followed by hydroxylamine cleavage to profile S-palmitoylated proteins. This confirmed 282 hydroxylamine-sensitive proteins from 501 putative palmitoylated proteins enriched from the initial 17-ODYA labeling. This also revealed and validated that palmitoylation of AMA1, a protein essential for host-cell invasion, is not required on invasion but increases microneme secretion. Taken together, these studies presented underscore the utility of large-scale S-palmitome profiling in understanding the biological importance of this lipid modification. Applying these techniques to future palmitome analysis would further discover potentially novel protein functions and cellular mechanisms across different biological systems. Wright, M. H., Heal, W. P., Mann, D. J. Tate, E. W. Protein myristoylation in health and disease. J. Chem. Biol. 3, 19-35 (2010). Farazi, T. A., Waksman, G. Gordon, J. I. The Biology and Enzymology of ProteinN-Myristoylation . J. Biol. Chem. 276 , 39501-39504 (2001). Heal, W. P., Wickramasinghe, S. R., Leatherbarrow, R. J. Tate, E. W. N-Myristoyl transferase-mediated protein labellingin vivo. Org. Biomol. Chem. 6, 2308-2315 (2008). Heal, W. P., Wright, M. H., Thinon, E. Tate, E. W. Multifunctional protein labeling via enzymatic N-terminal tagging and elaboration by click chemistry. Nat. Protoc. 7,105-117 (2012). Broncel, M. et al. Myristoylation profiling in human cells and zebrafish. Data Br. 4, 379-383 (2015). Charron, G. et al. Robust Fluorescent Detection of Protein Fatty-Acylation with Chemical Reporters. J. Am. Chem. Soc. 131, 4967-4975 (2009). Liu, Z. et al. Integrative Chemical Biology Approaches for Identification and Characterization of Erasers for Fatty-Acid-Acylated Lysine Residues within Proteins. Angew. Chemie Int. Ed. 54, 1149-1152 (2015). Wilson, J. P., Raghavan, A. S., Yang, Y.-Y., Charron, G. Hang, H. C. Proteomic Analysis of Fatty-acylated Proteins in Mammalian Cells with Chemical Reporters Reveals S-Acylation of Histone H3 Variants. Mol. Cell. Proteomics 10, M110.001198 (2011). Wright, M. H. et al. Validation of N-myristoyltransferase as an antimalarial drug target using an integrated chemical biology approach. Nat Chem 6, 112-121 (2014). Wright, M. H. et al. Global Analysis of Protein N-Myristoylation and Exploration of N-Myristoyltransferase as a Drug Target in the Neglected Human Pathogen Leishmania donovani. Chem. Biol. 22, 342-354 (2015). Wright, M. H., Paape, D., Price, H. P., Smith, D. F. Tate, E. W. Global Profiling and Inhibition of Protein Lipidation in Vector and Host Stages of the Sleeping Sickness Parasite Trypanosoma brucei. ACS Infect. Dis. 2, 427-441 (2016). Roberts, A. J. Fairlamb, A. H. The N-myristoylome of Trypanosoma cruzi. Sci. Rep. 6,31078 (2016). Thinon, E. et al. Global profiling of co- and post-translationally N-myristoylated proteomes in human cells. Nat Commun 5, (2014). Serwa, R. A., Abaitua, F., Krause, E., Tate, E. W. OHare, P. Systems Analysis of Protein Fatty Acylation in Herpes Simplex Virus-Infected Cells Using Chemical Proteomics. Chem. Biol. 22, 1008-1017 (2015). Burnaevskiy, N., Peng, T., Reddick, L. E., Hang, H. C. Alto, N. M. Myristoylome profiling reveals a concerted mechanism of ARF GTPase deacylation by the bacterial protease IpaJ. Mol. Cell 58, 110-122 (2015). Schmidt, M. F. G. Schlesinger, M. J. Fatty acid binding to vesicular stomatitis virus glycoprotein: a new type of post-translational modification of the viral glycoprotein. Cell 17, 813-819 (1979). Linder, M. E. Deschenes, R. J. Palmitoylation: policing protein stability and traffic. Nat Rev Mol Cell Biol 8, 74-84 (2007). Smotrys, J. E. Linder, M. E. Palmitoylation of Intracellular Signaling Proteins: Regulation and Function. Annu. Rev. Biochem. 73, 559-587 (2004). Martin, B. R. Cravatt, B. F. Large-scale profiling of protein palmitoylation in mammalian cells. Nat Meth 6, 135-138 (2009). Yount, J. S. et al. Palmitoylome profiling reveals S-palmitoylation-dependent antiviral activity of IFITM3. Nat Chem Biol 6, 610-614 (2010). Chesarino, N. M. et al. Chemoproteomics reveals Toll-like receptor fatty acylation. BMC Biol. 12,91 (2014). Santiago-Tirado, F. H., Peng, T., Yang, M., Hang, H. C. Doering, T. L. A Single Protein S-acyl Transferase Acts through Diverse Substrates to Determine Cryptococcal Morphology, Stress Tolerance, and Pathogenic Outcome. PLoS Pathog. 11,e1004908 (2015). Martin, B. R., Wang, C., Adibekian, A., Tully, S. E. Cravatt, B. F. Global profiling of dynamic protein palmitoylation. Nat Meth 9, 84-89 (2012). Roth, A. F. et al. Global Analysis of Protein Palmitoylation in Yeast. Cell 125, 1003- 1013 (2006). Kang, R. et al. Neural palmitoyl-proteomics reveals dynamic synaptic palmitoylation. Nature 456, 904-909 (2008). Morrison, E. et al. Quantitative analysis of the human T cell palmitome. Sci. Rep. 5, 11598 (2015). Srivastava, V., Weber, J. R., Malm, E., Fouke, B. W. Bulone, V. Proteomic Analysis of a Poplar Cell Suspension Culture Suggests a Major Role of Protein S-Acylation in Diverse Cellular Processes. Front. Plant Sci. 7, 477 (2016). Jones, M. L., Collins, M. O., Goulding, D., Choudhary, J. S. Rayner, J. C. Analysis of Protein Palmitoylation Reveals a Pervasive Role in Plasmodium Development and Pathogenesis. Cell Host Microbe 12, 246-258 (2012). Hernandez, J. L. et al. Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition. Mol. Biosyst. 12, 1799-1808 (2016). Zou, C. et al. Acyl-CoA:Lysophosphatidylcholine Acyltransferase I (Lpcat1) Catalyzes Histone Protein O-Palmitoylation to Regulate mRNA Synthesis. J. Biol. Chem. 286 ,28019-28025 (2011). Foe, I. T. et al. Global analysis of palmitoylated proteins in Toxoplasma gondii. Cell Host Microbe 18,501-511 (2015).

Natural and Sexual Selection of Vestigial and Wild Type Drosophila Melanogaster

Natural and Sexual Selection of Vestigial and Wild Type Drosophila melanogaster Abstract In this experiment, vestigial flies as well as wild type flies were used to create some diversity as well as test one of our hypotheses. Our hypotheses are as follows; 1) The wild type flies will have a greater relative fitness compared to the vestigial flies based on only sexual selection. The wild type has a relative fitness of 1 since it has fully functional wings. Vestigial Winged flies are not as sexually appealing since their wings are not functional and the wing movement is a vital part of their mating ritual. ) The vestigial fly will have a greater relative fitness when considering both sexual and natural selection. With the presence of natural selection (fly paper hanging from top of cage) posing a larger threat to the wild flies b/c they can fly better, this will allow vestigial flies to prosper better. To set up this experiment, two twenty-five gallon aquariums, 3 petri-dishes, 200 fli es, rotten bananas, and yeast were used. It was decided that there would be 80 vestigial flies and 20 wild type flies to total to an initial population of 100 drosophila.In the sexual selection circumstance, the vestigial drosophila’s carrying capacity is somewhere around 80 individuals, which is consistent with the wild type drosophila’s carrying capacity in this circumstance (Figure 4). In the sexual and natural selection circumstance the wild type drosophila have a carrying capacity at approximately 100 individuals (Figure 5). As both of the p-values from each circumstances in our experiment are greater than 0. 05, both hypotheses must be refuted. IntroductionThis experiment focuses specifically on Drosophila melanogaster, also known as the common fruit fly (Lynch, M. , et al. pp 645-663). For over 100 years scientists have been using them in experiments as they are referred to as a â€Å"model organism. † They were deemed a â€Å"model organism† since they are characterized as having a large reproductive capacity, a medium size, a short generation time, and are inexpensive to keep alive. Fruit flies only consume yeast growing on rotting fruit. Sixty-one percent of human genetic diseases have been found in fruit flies, they also share many similar genes with humans.The male fruit fly spends most of his time chasing after and singing to female flies with hopes of mating with her, they are successful about once a day (Stowe, K. 2008). The male fruit fly frequently sticks out one wing or the other which is accompanied by its vibration, producing a â€Å"love song† that can be recorded with specialized microphones (Hall, Jeffrey C. , pp 1702-1714). To distinguish male drosophila from females, there must be a distinctive black spot on the distal end of their abdomen for it to be a male, the females do not have this black spot.In this experiment, vestigial flies as well as wild type flies were used to create some diversity as we ll as test one of our hypotheses. Vestigial flies do not have well functioning wings, however wild flies do have functioning wings. Within this experiment, degrees of natural selection, sexual selection, and a combination of both were tested on the two types of flies. Natural selection is survival of the fittest. Sexual selection is the ability of organisms to choose their mates depending on their species. Relative fitness is the ability of an organism and species to survive and produce fertile, successful offspring.Purpose of our experiment is to determine the effect of natural selection on two subspecies coexisting under similar circumstances. Our hypotheses are as follows; 1) the wild type flies will have a greater relative fitness compared to the vestigial flies based on only sexual selection. The wild type has a relative fitness of 1 since it has fully functional wings. Vestigial Winged flies are not as sexually appealing since their wings are not functional and the wing moveme nt is a vital part of their mating ritual. 2) The vestigial fly will have a greater relative fitness when considering both sexual and natural selection.With the presence of natural selection (fly paper hanging from top of cage) posing a larger threat to the wild flies b/c they can fly better, this will allow vestigial flies to prosper better. Materials and Methods To set up this experiment, two twenty-five gallon aquariums, 3 petri-dishes, 200 flies, rotten bananas, and yeast were used. The bananas chosen to be an accelerant for the growth of the yeast and  were frozen so they would be easier to cut. The yeast was used because the drosophila melanogaster prefer this as a food source.The vestigial and wild type flies were sexed (to determine their sex), sorted, and counted. An initial population size of 100 total flies was decided so that it would be easier to determine the phenotypic percentage of the total population. Fly paper was placed in one of the sets of cages to impose a m ethod of natural selection as well as the sexual selection which is being solely tested by the other set of cages. It was decided that there would be 80 vestigial flies and 20 wild type flies to total to an initial population of 100 drosophila.Next, the flies were anesthetized flies using Fly Nap. The flies were counted out to reach desired ratio, sexing the flies making sure there are equal amounts of males and females to be sure there is ample individuals to allow successful mating. The fly’s food was prepared by taking a frozen rotten banana, cutting it in half, mashing up the banana meat, and mixing yeast into it. The bananas were then cut into halves horizontally (six halves total in each cage) and split down the middle length wise leaving the peel on the banana, revealing the yeast-banana mush and placed into the petri dishes.The petri dishes were then arranged into a horseshoe shape in the bottom of the aquarium. Next, the allele frequencies were determined by using th e hardy Weinberg equation (p2+2pq+q2). This experiment will prove our hypotheses sufficiently if carried out correctly by changing the total populations of the subspecies of each fly. Since the vestigial flies do not fly they will be affected by sexual selection only. However, the wild type flies will be affected by sexual and natural selection.The outcome that is predicted is that the relative fitness of the vestigial flies will stay low in the sexual selection circumstance and they will prosper in the sexual and natural selection circumstance since they do not fly and are not able to come near the fly paper posing as the natural selection. Also, the wild type flies will remain at a high fitness level in the sexual selection, but a low fitness level in both sexual and natural selection circumstance. Table 1: General Data from Sexual Selection| Generation| Sexual Selection Only| Total Drosophila| Wild Type| Vestigial| | 0| 20| 80| 100| 1|   |   |   | 2| 6| 12| 18| 3| 338| 50| 388| 4| 201| 21| 222| 5| 428| 35| 463| table 1: shown in this table is the initial data of the total count of flies captured for counting in each species each week/generation in the sexual selection group. | Results Table 2: General Data from Natural and Sexual Selection| Generation| Natural and Sexual Selection| Total Drosophila| Trapped Wild Type Flies| | Wild Type| Vestigial| | | 0| 20| 80| 100| 0| 1|   |   |   |   | 2| 16| 59| 75| lots| 3| 23| 10| 33| lots| | 86| 1| 87| lots| 5| 114| 21| 135| Lots| table 2: shown in this table is the initial data of the total count of flies captured for counting in each species each week/generation in the group that was exposed to natural selection as well as sexual selection. | Table 3: Anova: Single Factor, natural selection |   |   | Source of Variation| SS| df| MS| F| P-value| F crit| Between Groups| 63202. 5| 1| 63202. 5| 3. 513094| 0. 09775| 5. 317655| Within Groups| 143924. 4| 8| 17990. 55| | | | | | | | | | | Total| 207126. 9 | 9|   |   |   |   | | | | | | | |Table 3: Shown in this table is the ANOVA calculation which determines the probability that our null hypothesis will be found true. In this case, the proability of that happening is approximately 0. 09 or 9%. This number is low enough to allow us to accept our hypothesis. | Table 4: Anova: Single Factor natural and sexual selection|   | Source of Variation| SS| df| MS| F| P-value| F crit| Between Groups| 774. 4| 1| 774. 4| 0. 486447| 0. 505277| 5. 317655| Within Groups| 12735. 6| 8| 1591. 95| | | | | | | | | | | Total| 13510| 9|   |   |   |   | | | | | | | Table 4: Shown in this table is the ANOVA calculation for the Natural and Sexual selection grouping of drosophila and the possibility of our null hypothesis occurring. In this case, the value is approximately 0. 5, or 50%. This proves that our hypothesis is not completely reliable. | In â€Å"generation 0,† for the sexual and natural selection set of flies, the total sampl e size is 100 flies consisting of 20 wild type (10 male, 10 female), and 80 vestigial (40 male, 40 female), there were no flies stuck on the fly paper yet.The allelic frequency: p=0. 048, p= 0. 961. For the sexual selection set of flies, the total sample size is 100 flies, consisting of 20 wild type (10 male, 10 female), and 80 vestigial (40 male, 40 female), the allelic frequency is p= 0. 78, and q=0. 22. The dominant trait is denoted by the letter â€Å"p† (wild type), and â€Å"q† denotes the recessive (vestigial). After five generations were allowed to pass, the final total of drosophila in the sexual selection group included 428 wild type and 35 vestigial out of 463 total drosophila(Table 1).After five generations were allowed to pass, the final total of drosophila in the sexual and natural selection group included 114 wild type and 21 vestigial out of 135 total drosophila (Table 2). There is an obvious and drastic decrease in the amount of flies after the five ge nerations passed when comparing the two separate groups. Also, it must be pointed out that the set of data does not have values for the first generation because when the flies were counted, the collection of flies to count was unsuccessful.Next, the p-values of each set of data were calculated by using the ‘ANOVA: Single factor’ function in Microsoft Excel. The p-value of the sexual selection only set of data came out to be 0. 097, or 9. 7% that our null hypothesis that the vestigial flies would be more fit than the wild type flies would come true (Table 3). In Table 4, it is shown that the calculated p-value for the sexual and natural selection conditions of the vestigial and wild type flies produced a value of 0. 1, or 51% that our null hypothesis would come true. Figure 2 shows the data collected in the sexual selection circumstance of both the vestigial and wild type drosophila as well as the total population as to compare visually the drastic contrast between the t wo subspecies. The wild type flies remain more successful than the vestigial flies. Figure 1: In this figure, it is shown visually the differences in the numbers of flies counted per each subspecies of drosophila while under only sexual selection conditions.Figure 3 shows the Natural and sexual selection circumstance’s effects on the populations of both vestigial and wild type flies as well as the total number of flies in the enclosure. The Vestigial flies start off more prosperous than the wild type flies, but then the wild type drosophila regain their success and start reproducing at incredible rates. Figure 2: In this figure, it is shown visually the differences in the numbers of flies counted per each subspecies of drosophila while under both natural and sexual selection.Figure 4: In this figure, the relative fitness and carrying capacity of each subspecies is shown. Figures 4 and 5 have been included to compare fitness levels of each of the fly subspecies. The scale on t he y-axis has been multiplied by 10 to better understand the incredible contrast between the species. In both circumstances, the vestigial drosophila starts out with a higher relative fitness then reaches an equilibrium level with the wild type flies and they switch dominance of their relative fitness.Figure 5: In this figure, the relative fitness and carrying capacity of each subspecies is shown. Discussion This curvature of the graphs and intersections happen because when the subspecies reach their carrying capacities, they must make a change so that they can survive as a community. In the sexual selection circumstance, the vestigial drosophila’s carrying capacity is somewhere around 80 individuals, which is consistent with the wild type drosophila’s carrying capacity in this circumstance (Figure 4). In the sexual and natural selection ircumstance the wild type drosophila have a carrying capacity at approximately 100 individuals (Figure 5). As both of the p-values fr om each circumstances in our experiment are greater than 0. 05, both hypotheses must be refuted. It is suggested that this experiment is conducted again using a better enclosure for the flies. During the semester, countless flies were flying around the lab which could account for the numbers of flies being inaccurate. There are possible errors in the sexing as well as the counting of the flies.Only a few flies crawled into the tubes with apple cider vinegar, and didn’t allow for every fly to be accounted for. The temperature of the lab was often quite cold, therefore this could account for some of the flies dying. As temperature was not a factor we were testing in this experiment, the temperature should be kept at a constant level. This experiment is important on a global level because of the closeness in the similarities of the genes between the drosophila and humans (Lynch, M. , et al. pp 645-663).This experiment shows us how, if humans were meant to survive strictly on nat ural and sexual selection, that mutations would play a part in the selection of mates. References Hall, Jeffrey C. Jun 1994. The Mating of a Fly. Science 264 No. 5166: pp 1702-1714. Lynch, M. , J. Blanchard, Houle, D. , T. Kibota, Shultz, S. , L. Vassilieva, Willis, J. Perspective: Spontaneous Deleterious Mutation. Evolution 53 No. 3: pp 645-663. Stowe, K. A. , L. L. Hester, and Vieyra, M. L. 2008. Biology 101 Lab Manual. Hayden- McNeil Publishing Inc. , Plymouth, Michigan, USA.

A Shipwreck of Faith - Free Essay Example

Sample details Pages: 5 Words: 1580 Downloads: 3 Date added: 2019/03/21 Category Religion Essay Level High school Tags: Faith Essay Did you like this example? In Daniel Defoes book Robinson Crusoe, the main character endures many battles along his adventures while entering adulthood. As Robinson Crusoe journeys across the seas, God makes Crusoe enduress many trials. First, starting from when he sets off to sea, disobeying his parents wishes in the process, to encountering many shipwrecks, and lastly the opportunity of sharing his growing faith with others. By Crusoe encountering these many obstacles, it goes just to show how having a weak foundation in faith can reveal many unwanted paths. Christianity plays a major role in this well-known piece, as this is the biggest obstacle Crusoe is struggling with right from the jump. Crusoes father had a life plan set for him ever since he was little. He wanted his son to become a great lawyer, as stated, my fatherdesigned me for the law(Defoe 9). This was not in Crusoes heart obviously, as it was set for the ocean. Crusoe completely ignores what his father was trying to accomplish for his son, and set sail for the great adventure that the seas had to offer, showing us our first Christianity reference. In the Bible, the story of the prodigal son is a story of how two brothers both get share of their fathers inheritance. The father warns them to be careful of how they use such money, but only one son listens, as the other ignores his father and adventures what the world has to offer (New International Version, Luke 15.11-13). Don’t waste time! Our writers will create an original "A Shipwreck of Faith" essay for you Create order Crusoe is unaware of his actions as he has not developed a deep and prolonging faith within the religion of Christianity itself. In the beginning, he is what most Christians would describe as a lukewarm Christian. What this means in simpler terms is that he knows what Christianity is and says he believes in the customs and aspects of the religion, but does not live his life according to what is being described within the Bible itself. With the current situation that Crusoe is found within the beginning of the story, this takes a toll on his future escapades as his weak mind and faith have him struggle in the battles soon to come. As time moves on, Crusoe soon finds himself shipwrecked with nowhere to go. He is captured by Turkish pirates, who make Crusoe and many others their slaves. Slavery was very popular back in the time of Robinson Crusoe, so it was not looked as a terrible matter as seen today. Still, for Crusoe, he was going through a lot mentally, as one went from slave owner to slave is a drastic change in roles. Seeing this happening, readers could suggest that this was an act of God humbling Crusoe, and showing how little he was compared to everything else that was surrounding him every day, that life could change so quickly in an instant. While captured, he meets another slave, Xury. Later when Crusoe and Xury are rescued by a Portuguese sea captain, Crusoe makes a deal with the captain, to set Xury free on only one condition: he converts to Christianity (Xury in Robinson Crusoe par. 3). With this deal set between the captain and Crusoe, this demonstrates Crusoes first attempt at trying to share his steady growing faith within meeting new people. As God humbled Crusoe, it brought to the surface a different kind of sailor, who wants to show others what life is actually all about. Defoe never went into anymore detail of what happened with Xury and his call to action towards a new religion found upon by both of the slaves, but this was a very large step at the development towards Crusoes journey in the religion. Growing up with his parents, especially his father, Crusoe had a well-established education, not only for the base education taught in most schools, but within the profession his father wanted for him. Stated in the book, my father and mother about their being so positively determined against what they knew my inclinations prompted me to (Defoe 6). The profession of being a lawyer is pretty rough, not only going through and living the job out in your everyday life, but finding the differentiations between everyday people and others who differ in social class. Lawyers have to find certain kinds of tactics, how to use them, and how to apply them to certain people who might think or do things contrasting from the normal way of everyone else. With this in mind, Crusoe was probably taught this by his father, which could help him with his developing faith by putting thoughts to action. As the old saying goes, actions speak louder than words. A big example of this could be seen when Crusoe and Friday are on the cannibalistic island together. As Crusoe rescued Friday from the cannibals, he takes Friday under his ownership and shows him the basics of living, his language, and soon Crusoes second conversion of faith in someone (Robinson Crusoe Theme par. 1).. As Crusoe teaches Friday, Friday soon starts to understand a lot of Crusoes customs, but about his relationship with God. (RobinsonCrusoe par. 33) While this representation goes on, it all ties back to that original saying. Crusoe might have taught Friday about Christianity, but he also did it in action as well, most of the time unintentionally. Friday was rescued by Crusoe, but in the text he was seen calling Crusoe names of power like master (Friday in Robinson Crusoe par. 1). With this being stated, it is being depicted that Friday is under authority or power of Crusoe, much like a slave. As a slave, Friday does whatever Crusoe orders him to. Friday soon gets into this mind set later on, and since he is Crusoes property he wants to follow everything that Crusoe does, including religion. Not only was the religion of Christianity being taught, but Friday wanted to learn more and develop these thoughts to show his master that he respects him who leheartedly. This could be seen as a big leap of faith, not only by Friday, but by Crusoe himself. He is starting to develop a sturdy relationship between him and God, by bringing others to know Him. This is big for Crusoe because it is showing signs of maturity and trust as he is growing through the time spent away from home and stranded on the island. As Robinson Crusoe escapes the island and finds his way back home, he soon discovers that his plantation has made him a reasonably good amount of money. After he obtains his money from the plantation in Brazil, he pays everyone who has cared for him, starting from the Portuguese captain who rescued him and Xury, to one of his widow friends and two sisters. As this is done in the story, this could look to be another religious aspect in Christianity. Tithing is a very important matter in most churches, as it is seen as giving back to the one who has given. This is seen often, as in Christianity it is believed that the love and sacrifice that has been given to the followers of Christ is unpayable, but giving what they have to help the church is portrayed as a temporary way of doing so. This could be compared to Crusoes side of the story, as many have helped him and others along the way of his countless journeys and explorations all without any kind of payment or return at the time. As h e receives his profit, he instantly pays back the Portuguese captain who rescued him from shipwreck. He then returns to England to then repay a widowed friend and his two sisters, another example of Crusoe giving back to the ones who gave to him in the beginning. With evidence from the text and representations that back the religious aspects of Christianity, there is no doubt that Crusoe was struggling with a lack in faith. What Crusoe went through, starting from the beginning seen defying his parents wishes, to encountering new experiences through troubled times, and finally ending it with building a strong foundation in the faith of God shows that it was a tough ride for him all around. Many would say Crusoe has seen it all, but of course he sets back sailing again at the end of the story, adventuring even further to see what more the world has to offer. It is clearly shown in the story that the theme is constant all the way towards the end; No matter how tough the obstacles will become, God is always the guide towards a strong outcome. Crusoe constantly tried and tried again after every attempt to do things by himself, but as soon as he let God take the reins everything started to look brighter again in Crusoes future. As we face troubles and hardships, Crusoe should be a reminder to all that without a strong foundation in faith, it will lead us into many unwanted paths that for some, may be too difficult to endure. Works Cited Defoe, Daniel. Robinson Crusoe. 1719. Print. Shmoop Editorial Team. Robinson Crusoe Theme of Religion. Shmoop, Shmoop University, 11 Nov. 2008, www.shmoop.com/robinson-crusoe/religion-theme.html. RobinsonCrusoe A Born-Again Christian?, www.wordbasedcounseling.org/Articles/RobinsonCrusoe.htm. Shmoop Editorial Team. Xury in Robinson Crusoe. Shmoop, Shmoop University, 11 Nov. 2008, www.shmoop.com/robinson-crusoe/xury.html. Shmoop Editorial Team. Friday in Robinson Crusoe. Shmoop, Shmoop University, 11 Nov. 2008, www.shmoop.com/robinson-crusoe/friday.html. New International Version. Biblica, 2011. BibleGateway.com, www.biblegateway.com/versions/New-International-Version-NIV-Bible/

The Crisis Of The Subprime Loan Scandal - 1222 Words

The overarching problem of profiteering is the foundation of many of the moral problems that came from the exploitation of homeowners during the subprime loan scandal. In the financial industry, Lewis (2010) defines the awareness of top executives and bank loan officers that participated in giving out these loans, More so, an interview with Steve Eisman reaffirms the fully conscious role that loan officers played in allowing individuals without good credit ratings to take these loans. In many cases, the banking industry argued that it was the fault of those that took the loans, but it was actually the facilitation of these loans by loan officers that laid the unethical and immoral foundation of this scandal: Whenever Wall Street people†¦show more content†¦Another aspect of Desmond’s (2016)† research is based on the perspective of the slumlord culture that also exploits low-income people in the city of Milwaukee. A focus on the behaviors of local landlords also exposes the lack of social responsibility on the part of predatory property owners that only seek to extract money from the most impoverished sectors of society. Another aspect of the unregulated financial markets reveals the predatory nature of landlord policies, specially in the case of Tobin in Desmond’s (2016) sociological examination of real estate culture in urban Milwaukee. On one hand, Tobin would be lenient with renters in his trailer park when they were temporally unable to pay rent, but on the other hand, a lack of written contracts also defines the underhanded method of manipulation and control that he wielded over his renters. Much like the banking institution loan officers that never che cked the credit background of a subprime loan recipient, so does Tobin ignore contractual and legal obligations when managing his tenants in the trailer park: Tobin’s negotiations with tenants were rarely committed to writing, and sometimes tenants remembered things differently from the way Tobin did. A tenant would say she owed $150 and Tobin would say it was $250 or $600. Tobin once forgot tht a tenant paid a year’s worth of rent in advance after winning a worker’s compensation claim. Trailer park residents had a word for this: being â€Å"Tobined.† InShow MoreRelatedThe Subprime Loan Crisis : An Analysis Of The Ethical Shortcomings1495 Words   |  6 PagesThe Subprime Loan Crisis An Analysis of the Ethical Shortcomings Ten years ago the US housing market was booming and with a constant rise in prices there didn’t seem to be an end in sight. 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