Errors in information transfer from DNA to RNA to protein are inevitable. Transcription errors occur at a rate of ~10-5 per residue in Escherichia coli, over 10,000 times higher than errors in DNA synthesis. Errors in DNA synthesis can produce heritable changes in phenotype due to alteration of protein function. Studies that have focused on the mechanisms of DNA replication and repair in E. coli have provided a major framework for understanding the fidelity of genetic transmission from cell to cell and have revealed a series of fidelity mechanisms responsible for maintaining DNA integrity. Transcription errors, although transient in nature, can also have phenotypic consequences for the cell, including transient and heritable phenotypic change. Unlike DNA fidelity, the mechanisms ensuring transcription fidelity in vivo are not well characterized due to the difficulty of isolatig such transient errors in mRNA. The overall goal of this research is to (1) define the origins and the consequences of transcription errors in Escherichia coli, and (2) study the conflict between transcription fidelity and DNA repair. We have developed novel genetic tools to capture transcription errors and study their effect at the cellular level. Using next generation sequencing we will identify the spectrum of transcription errors and monitor how transcription fidelity hinder DNA break repair. This work will illuminate the fundamental cell/molecular biology of transcription fidelity in living cells, which are likely to be critical to many fundamental aspectsof biology and medicine including cancer, aging, and the resistance to drugs such as antibiotics.

Public Health Relevance

To accomplish their functions, cells constantly decode the information from their DNA. Errors in the transfer of information from DNA to proteins can trigger cellular dysfunction. Our study aims to understand the origin and consequences of these errors on cellular behavior.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM088653-09
Application #
9627994
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Sledjeski, Darren D
Project Start
2010-08-01
Project End
2020-01-31
Budget Start
2019-02-01
Budget End
2020-01-31
Support Year
9
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Genetics
Type
Schools of Medicine
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030
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Girard, Mary E; Gopalkrishnan, Saumya; Grace, Elicia D et al. (2017) DksA and ppGpp Regulate the ?S Stress Response by Activating Promoters for the Small RNA DsrA and the Anti-Adapter Protein IraP. J Bacteriol :
Sivaramakrishnan, Priya; SepĂșlveda, Leonardo A; Halliday, Jennifer A et al. (2017) The transcription fidelity factor GreA impedes DNA break repair. Nature 550:214-218
Han, Bing; Sivaramakrishnan, Priya; Lin, Chih-Chun J et al. (2017) Microbial Genetic Composition Tunes Host Longevity. Cell 169:1249-1262.e13
Xia, Jun; Chen, Li-Tzu; Mei, Qian et al. (2016) Holliday junction trap shows how cells use recombination and a junction-guardian role of RecQ helicase. Sci Adv 2:e1601605
Marciano, David C; Lua, Rhonald C; Herman, Christophe et al. (2016) Cooperativity of Negative Autoregulation Confers Increased Mutational Robustness. Phys Rev Lett 116:258104
Grace, Elicia D; Gopalkrishnan, Saumya; Girard, Mary E et al. (2015) Activation of the ?E-dependent stress pathway by conjugative TraR may anticipate conjugational stress. J Bacteriol 197:924-31

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