# Functional and structural studies of the transition from transcription initiation to elongation

> **NIH NIH R01** · UNIVERSITY OF PENNSYLVANIA · 2020 · $306,757

## Abstract

The goal of this proposal to determine the mechanisms of pol II transitions between
initiation, elongation, and re-initiation by pol II, which we will accomplish through
structural and biochemical means using the powerful Saccharomyces cerevisiae model
system. The in vitro transcription system we have developed for structural analysis is
based on stoichiometrically assembled, highly active and homogeneous components
comprising over 30 polypeptides, including RNA polymerase II (polII) and the general
transcription factors (GTFs). Critically, we have recently succeeded in optimizing this in
vitro reconstituted transcription system such that it achieves melting of double-stranded
promoter DNA, and initiation of RNA synthesis de novo with ~100% efficiency. We will
leverage this system to isolate and dissect, both structurally and biochemically a series
of pol II complexes representing a range of states, from initiation, to early elongation, to
complexes that might function as scaffolds for reinitiation. We will achieve these aims
through Cryo-EM, cross-linking and mass spectrometry (XL-MS), and quantitative XL-
MS.

## Key facts

- **NIH application ID:** 9982341
- **Project number:** 5R01GM123233-04
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** Kenji Murakami
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $306,757
- **Award type:** 5
- **Project period:** 2017-09-15 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9982341

## Citation

> US National Institutes of Health, RePORTER application 9982341, Functional and structural studies of the transition from transcription initiation to elongation (5R01GM123233-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9982341. Licensed CC0.

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