E2f Isoforms Lab Report

Introduction:

This experiment was preformed with the intent of discovering the affect of alcohol on E2F transcription factor proteins from HT-1080 cells. Western analysis was used to determine if alcohol has an effect on E2F isoforms.

The E2F transcription factor family has many important roles within the body. In general, the E2F family is important for controlling cell cycle and tumor suppression. (e2f1 e2f transcription factor 1). E2F transcription factors also contain several different domains relating to DNA binding, dimerization, transactivation, and tumor suppression. In total, there are eight isoforms of E2F transcription factor protein. The first isoform is E2F1, which has a molecular weight of approximately 46.9kDa (E2f gene protein

These three isoforms have a cyclin A binding domain, DNA binding domain, dimerization domain, and a transcriptional activation domain with a pocket protein binding domain. They are also all activators involved in G1/S cell cycle transactions. When these isoforms experience a loss of function it results in cell cycle arrest and when they are over expressed the cell cycle is driven into S phase (Attowooll). E2F4 and E2F5 isoforms have similar characteristics with molecular weights of approximately 44.0 kDa and 37.6 kDa respectively (E2f gene protein coding). These two isoforms have a DNA binding domain, dimerization domain, and a transcriptional activation domain with a pocket protein binding domain. Both isoforms are inhibitors that mainly work within nuclear G0/G1 cells and can bind to retinoblastoma proteins (pRB) (Attowool). The main function of these isoforms is to induce cell cycle exit and differentiation. If these isoforms are overexpressed they do not drive cells into the S phase. E2F6 has a molecular weight of

It is known that alcohol can cause damage and functional problems to protein, carbohydrates, and fat metabolism (mezzo). Due to this knowledge, it was expected that E2F transcription factor proteins would be affected by alcohol. In order to perform this experiment, HT-1080 cells were acquired from a fibrosarcoma of a 35-year-old human male. The concentration of protein in the HT-1080 cells was determined and then Western analysis was performed. Image J software was used to analyze the Western Blot in order to determine protein density and determine if there was a significant change of E2F isoforms density when exposed to