# Determinants of elongation rate differences between B. subtilis and E. coli RNA polymerases

> **NIH NIH F32** · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · 2021 · $65,994

## Abstract

PROJECT SUMMARY/ABSTRACT
Across all life, gene expression depends upon the faithful and timely transcription of messenger RNAs by RNA
polymerase. The molecular details of RNA polymerase (RNAP) elongation have been determined for enzymes
from a few species, but it is unclear whether these examples are representative of the whole. Recently, RNAP
from the model gram-positive bacterium Bacillus subtilis was found to be uncoupled from translation and travel
at a much faster rate than RNAP from its gram-negative counterpart, Escherichia coli. This observation is
complemented by known differences in transcriptional regulation between these species. I hypothesize that
because many mechanisms of gene regulation act upon elongating RNAP, changes in elongation rate will
affect how this regulation occurs. The goal of my proposal is to determine the causes of elongation rate
divergence between B. subtilis and E. coli and to examine the roles of B. subtilis RNAP-associated factors in
co-transcriptional gene regulation. Because pausing is a major determinant of RNAP elongation rate in vivo, in
Aim 1 of my proposal I will determine differences in pausing between B. subtilis and E. coli using nascent RNA
sequencing. In Aim 2, I will examine intrinsic and trans factors affecting B. subtilis and E. coli RNAP elongation
rate in vitro using single-molecule microscopy. In Aim 3, I will establish functions of B. subtilis RNAP-
associated factors with a genetic interaction screen. This work will illuminate the idiosyncrasies of B. subtilis
transcriptional machinery, which are likely shared by other Firmicutes bacteria including the pathogens S.
aureus and C. difficile. Because RNAP is an established antibiotic target, understanding differences in RNAP
function across bacteria could inform the development of novel species-specific antibiotics.
My fellowship training plan combines the knowledge and experience of my sponsors, Dr. Gene-Wei Li and Dr.
Jeff Gelles, to facilitate my research and career goals. Dr. Li has a strong background in quantitative biology
and will provide me training in the design of sequencing experiments and computational data analysis. Dr.
Gelles has years of experience in single-molecule biophysics that will be a major asset has I develop my own
skills in this area. Dr. Gelles has also mentored several post-doctoral trainees that have obtained independent
research positions. The support of my sponsors in combination with the research infrastructure and career
opportunities at the Massachusetts Institute of Technology will facilitate the completion of the research aims in
this proposal and my development as an independent scientist.

## Key facts

- **NIH application ID:** 10313703
- **Project number:** 1F32GM143898-01
- **Recipient organization:** MASSACHUSETTS INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** Robert A Battaglia
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $65,994
- **Award type:** 1
- **Project period:** 2021-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10313703

## Citation

> US National Institutes of Health, RePORTER application 10313703, Determinants of elongation rate differences between B. subtilis and E. coli RNA polymerases (1F32GM143898-01). Retrieved via AI Analytics 2026-05-29 from https://api.ai-analytics.org/grant/nih/10313703. Licensed CC0.

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