P53 Protein Lab Report

The purpose of this research was to determine the mechanisms in naked mole-rats that restricts cell proliferation and tumor formation. Contact inhibition is a mechanism observed where cell proliferation stops when two cells come in contact with one another. This process is caused by elevated levels of p27 cyclin-dependent kinase inhibitor which stops cells in the G1 phase from dividing. The cylcin-dependent kinase inhibitor p16 is also thought to regulate contact inhibition but its levels do not change in mice or human samples. Tumor-suppressor genes Rb and p53 are present in high concentration during contact inhibition to help control cell proliferation and apoptosis respectively. It was hypothesized that the naked mole-rats' cancer resistivity

| * is a common target of deletion or mutational inactivation in human tumors * an inhibitor of Cdk4-cyclin D complexes; * The loss of p16 leads to increased Cdk4-cyclin D activity

If the APC protein were to experience a nonsense mutation, the most common mutation in regards to this specific genetic disease, the destruction complex would not perform correctly and the wnt-signaling pathway would be stimulated. Problems begin to arise when “the Wnt/Beta-catenin pathway is activated [by] a Wnt ligand [binding] to seven-pass transmembrane Frizzled receptor and its co-receptor, low-density lipoprotein related protein 6 (LRP6) or its close relative LRP5” (MacDonald, et. al 2009). Once the wnt-signaling pathway is stimulated, a Wnt-Fz-LRP6 complex forms and takes in the Dishevelled (Dvl) protein, resulting in the phosphorylation and activation of the recruitment of the axin complex, destruction complex, to the receptors. (MacDonald, et al 2009). Due to the phosphorylation within the Wnt-Fz-LRP6 complex, the axin complex is not able to phosphorylate Beta-Catenin, which leads to the excessive accumulation of the protein in the cytoplasm. Researchers witnessed the development of numerous intestinal adenomas in transgenic mice with stabilized mutant Beta-catenin, therefore the results suggest that dysregulation of Beta-catenin is a key oncogenic event that follows the loss of APC function. (Tarapore, et. al 2011). The loss of the destruction complex’s normal activities results in elevated levels of Beta-catenin and downstream effectors such as CCND1,

The gene is located on chromosome seventeen (17p13.1). The genomic coordinates are 17:7,571,719-7,590,867 and it is 19,148 base pairs in length. The p53 protein is made up of three hundred and ninety three amino acids and is considered a phosphoprotein. The gene also contains four domains, also called units. Each one of these domains has a different responsibility. One domain, called the core domain, identifies specific DNA sequences. Another domain stimulates transcription factors. The third domain controls the tetramerization of the protein. The last domain is able to distinguish damaged DNA. This may include single-stranded DNA or base pairs that are misaligned (Vijayaraj).

Cancer is a name given to many diseases that are similar. In all kinds of cancer, blood cells divide uncontrollably(NCI). Science has had a huge affect on the way cancer is being diagnosed and treated. When doctors start to see symptoms like new moles/skin changes and quick weight gain or loss, they will do a number of tests, to see if you have cancer. Some of the tests include, Lab tests, biopsy, MRI, X-ray, and CT scan, though there are many additional tests. In lab tests, they check and see if the ratio of substances needed for your body are normal, or not. Normally, for these tests they use urine or blood. In a biopsy, a doctor will take a tissue sample and examine it to see if anything looks different. To remove tissue, doctors

One gene that is prominent in the regulation and development of cells is the Tumor Suppressor Gene. The function of the Tumor Suppressor Gene is to inhibit cell growth when it is deemed necessary in the body. When this gene is mutated is result in the overgrowth of cells because the body can not inform the proper cells to stop growing which can lead to cancer. Cells begin to take up space and oxygen without regard to the other cells around them. The gene “p53” is a common mutation of tumor suppressor genes and is said to be a cause and the development of cancers such as breast cancer. The problem with this mutation is that at the G1 checkpoint of interphase the mutated cell moves directly from G1 to S phase and produces two daughter cells with the mutated p53 gene. The Tumor Suppressor gene “apc” is also mutated in a majority of colorectal cancers. Researchers found that two protein inactivating mutations at apc result in sporadic tumors on the colerectum. Another gene that aids in cell reproduction is the Proto-Oncogene. The proto-oncogene stimulates cell growth. When this gene is mutated is results in the overgrowth of cells because the body is stimulating the cells to continue to divide and produce uncontrollably which can cause cancer because again cancer is defined as the uncontrollable division of cells. The oncogene version of the ras family has been proven to be present in a variety of cancers. The ras family has three major genes including H-ras, K-ras and N-ras. When ras genes are mutated they grow uncontrollably and evade death signals. The gene “k-ras” has played an early role in the development of pancreatic cancer. The last prominent gene that aid in cell cycle is DNA Repair Genes. DNA Repair Genes correct mistakes in DNA, mistakes which result in certain parts of the DNA producing the incorrect protein and function. When this gene is mutated the gene is

Biotechnology industry, Research, Pharmaceutical industry used purified proteins in large quantities for their purposes. This able genetic engineer to set up some technics to easily extracted from variable source proteins in large amount.

Embryonic stem cell research has led to medical benefits to aid in curing diseases and many cancer cases that have grown in today’s society. Tissues throughout the body have a specific single layer of cells that can regenerate daughter cells. These daughter cells, or embryonic cells, have the capabilities to regenerate and build tissues or organs. These cells can aid in a lifelong regeneration process for tissues throughout the body. In vitro studies have shown that these cells can be placed in a specific area where there is large amounts of tissue damage due to injury or disease and completely rebuild this tissue into a completely new, fully functioning tissue. (Weissman, 2005, p. 1). However, people will argue the fact that embryonic stem cells have not been cleared to work in the human body. This is true that Weisman has only found it to work in mice and in genetically made organs, tissues, and muscle. Facts have shown that

The overall analysis of this manuscript was indicating how a particular gene is connected to how tumors/cancer are suppressed and how tumors/cancer also grow and multiply. When the codon is manipulated and mutated, the point in which the codon becomes altered, or the point in which the p53 gene is stimulated, forms reactions that may be harmful or can be non-threatening. The reaction that could be harmful could generate destruction to the gene and therefore form cancer. A non-threatening reaction may cause a reconstruction in the mutation, which is beneficial.

The p53 protein has five highly sustained regions within a cross species amino acid sequence which are also known as domains. Of these five, there are four functional domains, transcriptional activation domain at the N-terminus,

The transcription factor p53 known as tumor suppressor gene, the main functions of P53 are activating the DNA repair, cell cycle arrest or induce the apoptosis pathway after DNA damage.

A missing or defective p53 protein is usually the culprit in cancer, being that is in over 50% of cancers. The p53 gene codes for a protein that stops the growth as well as the development of tumors, as well as other functions. This is known as a tumor suppressor gene. If the gene is mutated, it allows damaged cells to survive and make tumors. The p53 gene can be mutated by cancer-causing substances in the environment. The p53 is responsible for proteins that fix damaged cells or cause damaged cells to die (apoptosis). It does this so the damaged cells do not pass on. The p53 gene prevents irregular cells from turning into tumors.

Through the study, they did in fact find that while humans have 1 TP53 gene, African elephants have 19 different cancer suppressor genes. These genes were brought about through generations of evolution within the elephant community. It can be said that elephant cells demonstrated increased apoptotic response following DNA damage compared to human cells.

The technological advancements in embryonic stem cell research have been outstanding. In addition to the vaccines and treatments that scientists have already discovered, they hope to find more breakthroughs in the near future. (Xibo) Some of the research topics scientists are looking into is replacing damaged cells, finding defects in early development that can cause diseases or miscarriages, study normal development, and also learn how medicines or toxins affect a fetus. (Klimas) Embryonic stem cells are an incredible approach to medical research. These stem cells have no defined role in what they will become. They have the potential to turn into anything from brain cells or liver cells or even muscle cells. Embryonic stem cells divide rapidly and are also less susceptible to rejection than adult cells when transplanted, making embryonic stem cells the ideal source of stem cells. They can be transplanted into a place with some damaged cells and be able to replace them and work as if they were in place before the damage occurred. (Kobe)

Previous studies have suggested that K63 ubiquitination is associated with anti-apoptotic protein trafficking pathways. Figure 1 addresses whether K63 ubiquitination is accomplished by the TRAF6 E3 ligase. An immunoblot of p53 was used for U2SO cells in a Ni-NTA assay to determine if K63-linked ubiquitination of p53 was associated with TRAF6. U2SO cells were used because of their high proliferation rates. Flag-TRAF6 and p53 were included in the western blot as controls for this assay. Ubiquitin was tagged with histidine and TRAF6 was tagged with the Flag antibody to assess protein to protein interactions. This assay shows ubiquitination of p53 in the presence of His-Ub and Flag-TRAF6 indicating that K63 linked ubiquitination of p53 depends on TRAF6. To determine if specifically, K63-linked ubiquitination occurs through TRAF6 the authors assessed whether TRAF6 promotes K48-linked ubiquitination of p53 and proteasomal degradation. An immunoblot for HA in a Ni-NTA pull-down assay was generated to assess if ubiquitination and proteasomal degradation occurs at K48 and K63 by TRAF6. HA-p53 and Flag-TRAF6 in U2SO cells were used to show protein-protein interaction. A