# Project 3: Defining and targeting mechanisms of E2F transcription factor regulation

> **NIH NIH P01** · STANFORD UNIVERSITY · 2024 · $224,720

## Abstract

PROJECT SUMMARY
Activation of E2F transcription factors is the most downstream event in the retinoblastoma protein (Rb) tumor
suppressor pathway, which controls entry into the cell cycle and proliferation. E2F stimulates expression of
genes needed for DNA synthesis during S phase and for further progression into mitosis. Inactivation of Rb
and aberrant activation of the E2F-controlled transcription program is a hallmark of cancer. While Rb
repression of E2F has been extensively characterized, little is known regarding how E2F activates transcription
and how additional regulatory mechanisms control E2F function. Here we explore the structural mechanisms
underlying the function and posttranslational regulation of E2F.
Our first aim is to determine how E2F associates with chromatin and influences chromatin architecture to
induce gene expression. We will use structural, biochemical, and cellular assays to investigate how E2F binds
nucleosomes and how those interactions are critical for modulating chromatin architecture as cells activate the
transcription program for S phase entry. Our second aim is to identify novel E2F posttranslational modifications
and how modifications including phosphorylation and acetylation modulate E2F association with chromatin and
co-activator proteins. Our third aim is to reveal how E2F proteins are recognized by the E3 ligase adaptor
protein Cyclin F (CycF) for ubiquitylation and degradation. A structural and cell biology approach will be
applied to define the CycF-binding motif, and we will test the hypothesis that chemical inhibition of CycF can
deregulate the cell cycle in model cancer cells.
In collaboration with the other program projects and cores, our study will provide unprecedented molecular
analysis of E2F function and regulation. Our experimental results will provide foundational knowledge that can
be applied to design new therapeutic strategies that target E2F activity in diverse cancers.

## Key facts

- **NIH application ID:** 10809645
- **Project number:** 5P01CA254867-03
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Seth Michael Rubin
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $224,720
- **Award type:** 5
- **Project period:** 2022-03-25 → 2027-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10809645

## Citation

> US National Institutes of Health, RePORTER application 10809645, Project 3: Defining and targeting mechanisms of E2F transcription factor regulation (5P01CA254867-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10809645. Licensed CC0.

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