P53 Pathway Assay (CAT#: STEM-MB-0353-WXH)

Introduction

The p53 pathway is a key factor that helps to conserve the stability of the genome by preventing mutations caused by cellular stress or DNA damage. p53 stabilizes the genome by interacting with a number of signal transduction pathways in the cell to regulate a wide variety of cellular processes including apoptosis, senescence, cell cycle arrest, differentiation, and DNA repair and replication. p53 is activated in response to a number of stress stimuli, including DNA damage, oxidative stress and activated oncogenes, which are transmitted by post-translational modifications. Activated p53 functions effectively as a transcription factor and can then induce transcription of several genes. Mutations in p53 are associated with genomic instability and an increased susceptibility to cancer, particularly cervical and colorectal cancers, and it is believed that more than half of all cancers may involve p53-inactivating mutations. Additionally, certain DNA viruses, such as papillomavirus, can also inactivate the protein.

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Principle Applications Procedure Materials Notes Related Products

Principle

The p53 pathway responds to stresses that can disrupt the fidelity of DNA replication and cell division. A stress signal is transmitted to the p53 protein by post-translational modifications. This results in the activation of the p53 protein as a transcription factor that initiates a program of cell cycle arrest, cellular senescence or apoptosis. The transcriptional network of p53-responsive genes produces proteins that interact with a large number of other signal transduction pathways in the cell and a number of positive and negative autoregulatory feedback loops act upon the p53 response. There are at least seven negative and three positive feedback loops described here, and of these, six act through the MDM-2 protein to regulate p53 activity. The p53 circuit communicates with the Wnt-beta-catenin, IGF-1-AKT, Rb-E2F, p38 MAP kinase, cyclin-cdk, p14/19 ARF pathways and the cyclin G-PP2A, and p73 gene products. There are at least three different ubiquitin ligases that can regulate p53 in an autoregulatory manner: MDM-2, Cop-1 and Pirh-2.

Applications

To study the effect of each virus on p53 signaling pathway
To study the regulation mechanism of p53 signal pathway in disease
To study the effects of drugs or therapies on p53 signaling pathways

Procedure

• Luminex Multiplex Assay
• Enzyme-linked immunosorbent assay (ELISA)
• Flow cytometry (FACS analysis) technology

Notes

Detectable targets: ATM, ATR, CHK2, CHK1, MDM2, MDM-k, p14, p53, Cop-1, PIRH-2, CyclinG, Siah-1, Wip1, ANp73, TSAPS, PTEN, TSC2, IGF-BP3, p53R2, PAI, BAI-1, KAI, GD-Aif, TSP1, IGF, IGF-BP3, PIGs, Bax, PUMA, Siah, Fas, p53AIP, DRS, CASPS, Cdc2, CDK2, p21

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