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Human papillomavirus type 16 E7 regulates E2F and contributes to mitogenic signalling

J D Morris¹, T Crook, L R Bandara

¹Ludwig Institute for Cancer Research, St Mary's Hospital Medical School, London, UK.

|April 1, 1993

Summary

Human papillomavirus type 16 E7 protein can induce cellular DNA synthesis in mouse cells, especially with insulin. E7

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Area of Science:

Molecular Biology
Virology
Cell Biology

Background:

Human papillomavirus type 16 (HPV16) oncoproteins are implicated in cervical cancer.
The E7 protein is known to interact with cellular proteins to promote cell proliferation.

Purpose of the Study:

To investigate the mitogenic activity of HPV16 E7 protein in mouse fibroblasts.
To determine the role of E7 in inducing cellular DNA synthesis and its interaction with growth factors and transcription factors.

Main Methods:

Bacterial expression and purification of HPV16 E7 protein.
Scrape-loading assay to introduce E7 into quiescent Swiss 3T3 cells.
Assessment of cellular DNA synthesis and E2F transcription factor activation.

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Main Results:

HPV16 E7 protein alone weakly induced DNA synthesis in quiescent mouse fibroblasts.
E7's mitogenic activity was significantly enhanced by insulin.
E7 rapidly induced the active form of E2F, independent of insulin.
Mutant E7 proteins lacking RB binding capability failed to induce active E2F or synergize with insulin.

Conclusions:

E7's ability to regulate E2F is crucial for its mitogenic effects.
E7-mediated E2F regulation is necessary but not sufficient for full DNA synthesis induction.
Cooperation between E7 and growth factors like insulin is important for robust cell proliferation.