# The RNA polymerase II transcription complex

> **NIH NIH R01** · HARVARD MEDICAL SCHOOL · 2020 · $414,848

## Abstract

Project Summary. The goal of this project is to define the interactions between RNA polymerase II, the basal
and regulatory transcription factors, and the chromatin template that lead to accurate transcription initiation and
productive elongation. Using a combination of approaches in the yeast Saccharomyces cerevisiae model
system, several fundamental aspects of gene expression will be studied. Specific Aim 1 will continue our
studies of how transcription intersects with chromatin modifications. We will probe the function of the
Set1/COMPASS complex, which methylates histone H3 at lysine 4 (H3K4). Methylation is co-transcriptional,
and H3K4 trimethylation is strongest at the nucleosome nearest the transcription start site, while dimethylation
predominates further downstream through the next few nucleosomes. This aim will explore how
Set1/COMPASS is recruited to transcription complexes and how the gradient of methylation is created. In the
previous funding period we successfully developed a pipeline for proteomic analysis of both initiation and
elongation complexes using immobilized chromatin templates and quantitative mass spectrometry. Specific
Aim 2 will exploit these systems to further define the events and factors that drive the transcription cycle
forward. We also test a new model for how TFIID functions. In Specific Aim 3, our immobilized template
transcription system will be combined with single-molecule TIRF microscopy to visualize transcription in real
time. We will analyze the interaction kinetics between factors and the transcription template. Interesting
findings in all these aims will be validated in vivo using the genetic, genomic, and molecular techniques our lab
has developed over many years. The experiments in these three specific aims will significantly increase our
understanding of the RNA polymerase II transcription reaction and its interactions with the chromatin template.
Although the project uses a model system, mechanisms of transcription are highly conserved in eukaryotes
and, based on past experience, the results are likely to be directly applicable to human gene expression. This
fundamental knowledge is essential for understanding how mutations in transcription factors and histone
modifying enzymes lead to diseases such as cancer and developmental defects.

## Key facts

- **NIH application ID:** 9894809
- **Project number:** 5R01GM046498-29
- **Recipient organization:** HARVARD MEDICAL SCHOOL
- **Principal Investigator:** Stephen Buratowski
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $414,848
- **Award type:** 5
- **Project period:** 1991-07-06 → 2021-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9894809

## Citation

> US National Institutes of Health, RePORTER application 9894809, The RNA polymerase II transcription complex (5R01GM046498-29). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9894809. Licensed CC0.

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