All cellular organisms rely on a multi-protein molecular machine named the replisome to duplicate their genomes. At replication origins, assembly of replisomes for chromosomal DNA replication is a highly regulated event that is coordinated with the cell cycle. In Escherichia coli, DnaA protein initiates chromosomal DNA replication by orchestrating a step-wise process that leads to two DNA helicases bound to each parental DNA strand, which is necessary for bidirectional fork movement from the chromosomal origin. Other components of the replisome then assemble at the newly formed replication fork. Duplication of the genome follows. The recruitment of the replicative helicase to the chromosomal origin is a critical step during the initiation stage of DNA replication. The long-term objective of this research is to understand the molecular mechanism of recruitment and activation of DnaB, the replicative helicase of Escherichia coli, via genetic, biochemical and molecular biological methods. Because all free-living organisms use a similar mechanism of initiation, these studies should provide insight into how this event occurs in higher organisms.

Public Health Relevance

Chromosomal DNA replication is an essential process that occurs by similar biochemical mechanisms in all free-living organisms. At the replication origin of the Escherichia coli chromosome, a critical event is the loading and activation of the DNA helicase that functions to unwind the duplex DNA to propagate movement of the replication fork. This project investigates the molecular mechanism of recruitment and activation of DnaB helicase during the initiation of chromosomal replication;these studies may lead to novel methods to treat human diseases caused by bacterial pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM090063-02
Application #
8328635
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Reddy, Michael K
Project Start
2011-09-05
Project End
2015-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
2
Fiscal Year
2012
Total Cost
$258,093
Indirect Cost
$87,093
Name
Michigan State University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
193247145
City
East Lansing
State
MI
Country
United States
Zip Code
48824
Bell, Stephen P; Kaguni, Jon M (2013) Helicase loading at chromosomal origins of replication. Cold Spring Harb Perspect Biol 5:
Felczak, Magdalena M; Kaguni, Jon M (2012) The rcbA gene product reduces spontaneous and induced chromosome breaks in Escherichia coli. J Bacteriol 194:2152-64
Kaguni, Jon M (2011) Replication initiation at the Escherichia coli chromosomal origin. Curr Opin Chem Biol 15:606-13