P53 Protein Case Study

Its locus is particularly amplified in these noted tumours leading to the progression of these cancers, it can be suppressed by p53 (tumour/ proliferation suppressor) which represses the EZH2 promoter, resulting inhibition of cell proliferation and invasion (Bracken, 2003; Xiao, 2011).

To mutagenize human Kif5A (WT), we performed PCR-mediated site-directed mutagenesis to insert our mutation into the plasmid vector (shown to the right). Next, we performed a restriction enzyme digest with Dpn1, to digest the parental plasmid.  The presence of the PCR product was confirmed by performing gel electrophoresis where the expected band of 6.8 kilobases was obtained. The remaining DNA was used in a bacterial transformation of DH5α to grow colonies. Liquid cultures were grown from overnight colonies and Minipreps were performed to purify the plasmid DNA. These plasmids were sent for sequencing, where the P278L mutation was confirmed.

The maximum cell density of the naked mole rat cells was found to be three times lower than the mouse sample. This result shows that naked mole-rat cells are hypersensitive to contact inhibition, also known as early contact inhibition. Researchers attempted to determine whether this early contact inhibition was caused by cell contact or secreted factors by replacing the media of naked mole-rat fibroblasts. The replacement increased the maximum cell density but not enough to reach the same level as the mouse sample showing that contact is the cause of the contact inhibition. Naked mole-rat and mouse fibroblasts were infected with oncoproteins which disable Rb and p53 in different samples to determine their role in early contact inhibition. The results showed that both Rb and p53 both played a role in preventing cell proliferation but Rb is more important in the regulation of early contact inhibition. Naked mole-rat fibroblasts were compared to human, mouse and a mutated naked mole-rat fibroblast without early contact inhibition by analyzing for p27 using Western blot. The naked mole-rat sample was the only sample that expressed little p27. When the same process was repeated for p16, the naked mole-rat sample was the only sample expressing high levels of p16. This result shows how p16 is the early contact inhibitor in the naked mole rat and p27 serves as a backup. GFP

In the book p53: The Gene That Cracked The Cancer Code by Sue Armstrong, they focus on statistics rather than a story. This book describes the change throughout time while focusing on the p53 gene. It talks about different genes that affect p53 and how cancer can happen.

In the context of the cell cycle, P53 is shown to have a G1 and G2/M checkpoint function [23]; in fact, upon receiving a stimulus such as DNA damage, p53 induces cell cycle arrest providing time for the cell to repair the genomic damage before being released back into the proliferating pool . The best known P53 target gene product involved in this process is the cyclin-dependent kinase (CDK) inhibitor p21 [18]. The progression through the S phase of the cell cycle is tightly controlled by CDKs [19]. P21 functions by inhibiting Cyclin-CDK complexes, therefore, hindering the cell cycle transition from G1 to S phase [23]. In addition to being implicated in the G1/S arrest of the cell cycle, it has been demonstrated that P21, alongside p53, is also essential in the G2/M phase [23,

The concept of citizenship and its general approach in different eras by Michael Schudson has been such a topic for debate, given with the notion of how society today tend to define the distinct ideal “citizen” and its origins in America while comparing them to various nations around the world to see how each differs from the other while indicating that an informed citizen in a mass democracy has thus far been insufficient. He gave some examples in an earlier in an American democracy that an informed citizen did not in fact require much in society, with only traits such as loyalty and regularity being the most sought after; elections through education in an effort by “Mugwumps” (a term used to describe Republican political activists who left

In analyzing the effect of 2-aminopurine on mutation frequencies of bacterial strain CC102 (ara∆(gpt-lac)5 thi rpsL/F’ lacZ-Y+A+proA+B+ , the bacterial strain CC102 was cultured in LB medium with 2-aminopurine and one LB medium without 2-aminopurine. The bacterial strain CC102 with and without the 2-aminopurine mutagens were diluted to 10-5 and 10-6 and were plated on LB plates with nutrients and sources of carbon compounds for growth. The undiluted bacterial strain CC102 was then plated on minimal medium with thiamine and lactose. In noting the genotype of the bacterial strain, there is a deletion in the lac gene as it is ∆(gpt-lac) and the lacZ gene is not functional, therefore β-galactosidase is not able to cleave lactose into the constituents of glucose and galactose. In table 1.1, when observing the number of mutant colonies on minimal lactose and single cell colonies formed on LB plates with no introduction of the 2-AP mutagen, the mutant colonies were not formed on the minimal lactose yet were formed on the LB nutrient rich plate. For example in looking at group 6 data, 163 colonies were formed on LB medium at 10-6 yet no colonies were formed on minimal lactose with no 2-AP. Since the lac gene and the lacZ gene need to be functional to cleave lactose, it would allow for growth on the minimal medium with lactose but since there was a deletion for the lac gene and lacZ gene was not functional, no growth occurred. However with the introduction of the

5812). Since Alu elements are typically viewed as the cause of human mutations, they will have to be used as the testing tool (Yang et al. 5812). The DNA sequence from the mouse cells will be isolated so that it could be cloned into the bacteriophage (Sherif and Broome 124). Using the transposon tagging framework, which implies tagging the genes that do not coincide with the regular process of gene expression, one will be able to detect the presence of the human cellular oncogene in the fibroblasts developed by the mice (Zhang et al. 3134). The hybridization of a phage and the Alu probe will point to the presence of the human mutagen in the cell (Yang et al.

The ability of p53 to regulate metabolism is also associated with the ability to regulate cellular ROS levels. As previously mentioned, p53 can either remove damaged cells that have suffered sustained oxidative stress, or limit levels of ROS in order to lower oxidative stress and consequently, potential cell damage. Through the regulation of carbohydrate and lipid metabolism, p53 is able to influence the response to ROS accordingly. By driving the expression of TIGAR and promoting PPP activity, p53 can increase the production of NAPDH, which can be used to generate the cellular antioxidant GSH (Bensaad 2006). Moreover, at the expense of nucleotide synthesis, p53 can also promote GSH synthesis following serine starvation, thereby lowering ROS

Proteins are a very important part of the human body. Each of these proteins has a specific job in our body. One decides the color of hair, one decides the color of eyes, or one may keep the human body from diseases by making it immune. When some of these proteins are affected they will work in an inverse way causing harm to the human body. TP53 is an example of a protein that works in an uncontrolled way, possibly from the exposure of uranium and other materials, causing diseases such as cancer. In this paper, the radioactivity and the high chemical toxicity of uranium will be discussed. Also, when DNA and RNA become damaged, it then causes other proteins to

The purpose of this experiment was to determine whether a point mutation on the lac- strain, CSH61, from the previous experiment is suppressed by amber suppressors, SuI, SuII, and SuIII. If the point mutation on the lac- strain was a nonsense mutation, a second mutation on the tRNA for that codon would be able to restore function of the lac gene by inserting an appropriate amino acid at the stop codon instead of terminating protein production. Therefore, the CSH61 cells could grow on lactose. If the point mutation was missense, no growth would be able to occur as the lac gene would not have its function restored.

target misfolded protein degradation53. However, a major gap in the field is the lack of any

Beth has a few options to determine if her baby is at risk for having a genetic disorder. The first step Beth can take is visiting her gynecologist. While there she can start to discuss her concerns, and go over her options. She can go through genetic counselling and then move onto ultrasounds and amniocentesis. If further tests warrant she can do chorionic villus sampling or CVS.

~0 mV versus Normal Hydrogen Electrode (Rodgers and Sligar, 1991) suggests that electron transfer to ferric P450 (redox potential ~300 mV vs. NHE) is unfavorable. Hence it was suggested that the redox function of cyt b5 involved electron transfer to the ferrous dioxygen intermediate which has a redox potential near 0 mV (Lipscomb et al., 1976) thus providing the “second electron” in the normal monooxygenase stoichiometry. In an attempt to differentiate between these two roles, Coon and co-workers reconstituted apo cyt b5 with manganese protoporphyrin IX (Morgan and Coon, 1984). They found cytochrome P450 reductase (CPR) and NADPH could not reduce the manganese substituted cyt b5, whereas iron cyt b5 was rapidly reduced. Hence Mn b5 is incapable of any electron transfer to the P450. They concluded that cyt b5 effects depend on the specific P450 in question, the substrate being examined, and molar ratio of CPR to P450. This suggested that their observations could not be explained solely by a simple electron transfer role and some effects may also be caused by possible conformational changes caused by cyt b5 binding.

What would you do if, at no fault of your own, your entire life was turned upside-down and you were forced into an entirely different situation than the one you were previously in? In the novel, The Book Thief, by Markus Zusak, a young girl is put into tough situations, forcing her to make decisions that a girl her age would never even dream about having to make. During World War II, a young girl, Leisel Meminger, is forced to go live with a foster family in the town of Munich in Germany. Here, she learns of life, love, loss, and death. Many of her situations she is forced into doing, whether it is from her mama or the poverty that she are living in. In the small town of Munich, she makes friends, loses friends, and learns the pain of the world