1.

INTRODUCTION
The p53, also known as TP53 or tumor protein (EC :2.7.1.37) is a gene that codes for a
protein that regulates the cell cycle and hence functions as a tumor suppression. It is very important for
cells in multicellular organisms to suppress cancer. P53 has been described as "the guardian of the
genome", referring to its role in conserving stability by preventing genome mutation (Strachan and Read,
1999). The name is due to its molecular mass: it is in the 53 kilodalton fraction of cell proteins.
2.HISTORY
The p53 was identified in 1979 by Arnold Levine,David Lane and William Old,working at
Princeton University, Dundee University (UK) and Sloan-Kettering Memorial Hospital,
respectively. It had been hypothesized to exist before as the target of the SV40 virus, a strain that
induced development of tumors.Although it was initially presumed to be an oncogene, its character
as a tumor suppressor gene was revealed in 1989.In 1993, p53 protein has been voted molecule of
the year by the Science magazine
3. GENE
The human p53 gene is located on the seventeenth chromosome (17p13.1).
4. STRUCTURE
The p53 protein is a phosphoprotein made of 393 amino acids. It consists of four
units (or domains):

A domain that activates transcription factors.

A domain that recognizes specific DNA sequences (core domain).

A domain that is responsible for the tetramerization of the protein.

A domain that recognized damaged DNA, such as misaligned base pairs or single-stranded DNA.

Wild-type p53 is a labile protein, comprising folded and unstructured regions

which function in a synergistic manner (Bell et al. 2002).p53 protein has been voted
molecule of the year.
5. MECHANISM
It plays an important role in cell cycle control and apoptosis. Defective p53 could
allow abnormal cells to proliferate, resulting in cancer. As many as 50% of all human tumors
contain p53 mutants.
In normal cells, the p53 protein level is low. DNA damage and other stress signals
may trigger the increase of p53 proteins, which have three major functions:growth arrest, DNA

repair and apoptosis (cell death). The growth arrest stops the progression of cell cycle,
preventing replication of damaged DNA. During the growth arrest, p53 may activate the
transcription of proteins involved in DNA repair. Apoptosis is the "last resort" to avoid
proliferation of cells containing abnormal DNA.
Proteomics
STUDENT ACTIVITY
One regulatory protein implicated in causing cancer is p53. Go to the NCBI Web site andlook up the
protein sequence of human p53 (accession number AF307851) and mousep53 (accession number
X00741).
The NCBI Web site link is:
http://www.ncbi.nlm.nih.gov/
Then use the protein analysis toolkit (ExPASy proteomics toolkit page) to analyze thesetwo p53
proteins.
The ExPASy proteomics toolkit Web site link is:
http://us.expasy.org/tools/
Using the two Web sites, answer the following questions:
1. What kind of tools are available? List 5 different kinds and a briefdescription of what
each type does.
2. Use the ProtParam tool for both protein sequences. How many aminoacids does each
protein have? What is the theoretical molecularweight for each protein?
3. Use the Prosite tool for one of the proteins. What post-translationalmodifications are
possible on p53? Are these all actually found onthe protein inside the human or mouse?
4. Do a sequence alignment of the two proteins using the SIMalignment tool. What do the *
symbols mean below the aligned aminoacids? What is the percent identity? What does that
mean?
Hint: Begin your analysis using the Proteomics toolkit tools by cutting and pasting thep53 protein
sequences from the NCBI gene listing. Be careful to copy the proteinsequence and not the DNA
sequence.

The p53, also known as TP53 or tumor protein (EC :2.7.1.37) is a gene that codes for a
protein that regulates the cell cycle and hence functions as a tumor suppression. It is very important for
cells in multicellular organisms to suppress cancer. P53 has been described as "the guardian of the
genome", referring to its role in conserving stability by preventing genome mutation (Strachan and Read,
1999). The name is due to its molecular mass: it is in the 53 kilodalton fraction of cell proteins
Answer: