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The Possible Role of E2F in Rat Mammary Carcinogenesis

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Annual rept. 15 Jul 97-14 Jul 98

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My project, as described in the last annual report, was to coexpress E2F wild type and mutant proteins, ras, and GFP from the same viral vector and infuse the viruses into rat mammary glands to see if E2F wild type and mutant proteins could cooperate or interfere with ras mediated mammary carcinogenesis. While cloning of the viral vectors was successful and protein expression of the E2F proteins was evident, there was no ras expression from the IRES internal ribosome entry site as monitored by Westerns or NIH 3T3 transformation assays. Therefore, using viral infusion to investigate the interaction between ras and E2F proteins was not possible. As a result, I used tissue culture assays to investigate these two proteins in transformation. While E2F1 was were unable to cause transformation by itself, a E2F1 de1417-437 mutant interfered with ras transformation. Interestingly, this mutant does not affect cell cycle progression and does not appear to be toxic. Therefore, E2F1 de1417-437 may be affecting transformation-specific E2F targets. Endogenous E2F targets are being investigated by a chromatin immunoprecipitation assay.

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  • Anatomy and Physiology
  • Medicine and Medical Research

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