NDLI: Replication Initiation at a Distance

Content Provider	PubMed Central
Author	Saxena, Mukesh Abhyankar, Mayuresh Bastia, Deepak
Abstract	Plasmid R6K, which contains 3 replication origins called α, γ, and β, is a favorable system to investigate the molecular mechanism(s) of action at a distance, i.e. replication initiation at a considerable distance from the primary initiator protein binding sites (iterons). The centrally located γ origin contains 7 iterons that bind to the plasmid-encoded initiator protein, π. Ori α, located at a distance of ∼4 kb from γ, contains a single iteron that does not directly bind to π but is believed to access the protein by π-mediated α-γ iteron-iteron interaction that loops out the intervening ∼3.7 kb of DNA. Although the cis-acting components and the trans-acting proteins required for ori γ function have been analyzed in detail, such information was lacking for ori α. Here, we have identified both the sequence elements located at α and those at γ, that together promoted α activity. The data support the conclusion that besides the single iteron, a neighboring DNA primase recognition element called G site is essential for α-directed plasmid maintenance. Sequences preceding the iteron and immediately following the G site, although not absolutely necessary, appear to play a role in efficient plasmid maintenance. In addition, while both dnaA1 and dnaA2 boxes that bind to DnaA protein and are located at γ were essential for α activity, only dnaA2 was required for initiation at γ. Mutations in the AT-rich region of γ also abolished α function. These results are consistent with the interpretation that a protein-DNA complex consisting of π and DnaA forms at γ and activates α at a distance by DNA looping.
Related Links	http://dx.doi.org/10.1074/jbc.m109.067348
Ending Page	5712
Page Count	8
Starting Page	5705
File Format	PDF
ISSN	00219258
e-ISSN	1083351X
Journal	The Journal of Biological Chemistry
Issue Number	8
Volume Number	285
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	2010-02-19
Access Restriction	Open
Rights Holder	American Society for Biochemistry and Molecular Biology
Subject Keyword	Cell Biology Biochemistry Molecular Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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