Control of elongation of DNA replication Accurate and complete DNA replication is of paramount importance for all organisms. Replication elongation is highly susceptible to disruptions leading to fateful consequences including incomplete replication, mutations and genome rearrangements, which can underlie cell death, genomic disorders, tumor formation and progression. Despite the importance of minimizing disruption of DNA replication elongation, how cells manage to do so remains poorly understood. We recently employed genome-wide, systematic approaches to identify a novel nutrient-responsive regulation of replication elongation in the Gram-positive bacterium Bacillus subtilis. We propose that this regulation keeps replication elongation under metabolic control to prevent disruption of replication, and together with mechanisms that rescue and reactivate the disrupted replication forks, maintains genome integrity robustly. Such mechanisms are likely to exist in other organisms and play far more important roles than previously conceived.
We aim to evaluate the role of regulation of replication elongation, its prevalence in other bacteria such as E. coli, and to define the molecular differences between regulated replication arrests and disruptive replication arrests. Our study will contribute significantly to knowledge of the connection between the replication elongation complex and its cellular environment. This will deepen the understanding of the possible roles of uncontrolled replication in tumorigenesis and genomic disorders and elucidate fundamental principles underlying genome integrity.
The specific aims are: 1. Examine the role of a novel regulatory mechanism of replication elongation in B. subtilis;2. Characterize key events at replication forks following diverse replication arrests;3. Characterize the control of replication elongation in E. coli.

Public Health Relevance

We discovered a new way that bacteria control duplication of their DNA. Investigating this pathway elucidates novel and conserved principles of control of DNA duplication. Loss of this control potentially leads to genome rearrangements and mutagenesis, which underlies several human genomic disorders and cancers. Understand this pathway is also important for understanding microbial development of antibiotic resistance.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM084003-04
Application #
8269770
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Reddy, Michael K
Project Start
2009-05-01
Project End
2012-10-31
Budget Start
2012-05-01
Budget End
2012-10-31
Support Year
4
Fiscal Year
2012
Total Cost
$58,895
Indirect Cost
$4,950
Name
Baylor College of Medicine
Department
Genetics
Type
Schools of Medicine
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030
Li, Wenting; Bouveret, Emmanuelle; Zhang, Yan et al. (2016) Effects of amino acid starvation on RelA diffusive behavior in live Escherichia coli. Mol Microbiol 99:571-85
Gaca, Anthony O; Kudrin, Pavel; Colomer-Winter, Cristina et al. (2015) From (p)ppGpp to (pp)pGpp: Characterization of Regulatory Effects of pGpp Synthesized by the Small Alarmone Synthetase of Enterococcus faecalis. J Bacteriol 197:2908-19
Liu, Kuanqing; Bittner, Alycia N; Wang, Jue D (2015) Diversity in (p)ppGpp metabolism and effectors. Curr Opin Microbiol 24:72-9
Liu, Kuanqing; Myers, Angela R; Pisithkul, Tippapha et al. (2015) Molecular mechanism and evolution of guanylate kinase regulation by (p)ppGpp. Mol Cell 57:735-49
Zhang, Yan; Mooney, Rachel A; Grass, Jeffrey A et al. (2014) DksA guards elongating RNA polymerase against ribosome-stalling-induced arrest. Mol Cell 53:766-78
Kriel, Allison; Brinsmade, Shaun R; Tse, Jessica L et al. (2014) GTP dysregulation in Bacillus subtilis cells lacking (p)ppGpp results in phenotypic amino acid auxotrophy and failure to adapt to nutrient downshift and regulate biosynthesis genes. J Bacteriol 196:189-201
Maduike, Nkabuije Z; Tehranchi, Ashley K; Wang, Jue D et al. (2014) Replication of the Escherichia coli chromosome in RNase HI-deficient cells: multiple initiation regions and fork dynamics. Mol Microbiol 91:39-56
Bittner, Alycia N; Kriel, Allison; Wang, Jue D (2014) Lowering GTP level increases survival of amino acid starvation but slows growth rate for Bacillus subtilis cells lacking (p)ppGpp. J Bacteriol 196:2067-76
Arjes, Heidi A; Kriel, Allison; Sorto, Nohemy A et al. (2014) Failsafe mechanisms couple division and DNA replication in bacteria. Curr Biol 24:2149-55
Denapoli, Jessica; Tehranchi, Ashley K; Wang, Jue D (2013) Dose-dependent reduction of replication elongation rate by (p)ppGpp in Escherichia coli and Bacillus subtilis. Mol Microbiol 88:93-104

Showing the most recent 10 out of 17 publications