Intracellular Locations of Replication Proteins and the Origin of Replication during Chromosome Duplication in the Slowly Growing Human Pathogen Helicobacter pylori

We followed the position of the replication complex in the pathogenic bacterium Helicobacter pylori using antibodies raised against the single-stranded DNA binding protein (HpSSB) and the replicative helicase (HpDnaB). The position of the replication origin, oriC, was also localized in growing cells...

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Bibliographic Details
Published in:Journal of Bacteriology 2014-03, Vol.196 (5), p.999-1011
Main Authors: Sharma, Atul, Kamran, Mohammad, Verma, Vijay, Dasgupta, Santanu, Dhar, Suman Kumar
Format: Article
Language:English
Subjects:
antibodies
Bacteria
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Bacteriology
Cell Division - physiology
cell membranes
chromatids
Chromosomes
cohesion
DNA Replication - physiology
DNA, Bacterial - physiology
DNA-binding proteins
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
fluorescence in situ hybridization
Gene Expression Regulation, Bacterial - physiology
Helicobacter pylori
Helicobacter pylori - cytology
Helicobacter pylori - genetics
Helicobacter pylori - growth & development
Helicobacter pylori - metabolism
humans
pathogens
Protein Transport - physiology
Proteins
replication origin
single-stranded DNA
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Summary:We followed the position of the replication complex in the pathogenic bacterium Helicobacter pylori using antibodies raised against the single-stranded DNA binding protein (HpSSB) and the replicative helicase (HpDnaB). The position of the replication origin, oriC, was also localized in growing cells by fluorescence in situ hybridization (FISH) with fluorescence-labeled DNA sequences adjacent to the origin. The replisome assembled at oriC near one of the cell poles, and the two forks moved together toward the cell center as replication progressed in the growing cell. Termination and resolution of the forks occurred near midcell, on one side of the septal membrane. The duplicated copies of oriC did not separate until late in elongation, when the daughter chromosomes segregated into bilobed nucleoids, suggesting sister chromatid cohesion at or near the oriC region. Components of the replication machinery, viz., HpDnaB and HpDnaG (DNA primase), were found associated with the cell membrane. A model for the assembly and location of the H. pylori replication machinery during chromosomal duplication is presented.
ISSN:0021-9193
1098-5530
1098-5530
1067-8832
DOI:10.1128/JB.01198-13