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PMID: 1694129 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Strand separation required for initiation of replication at the chromosomal origin of E.coli is facilitated by a distant RNA--DNA hybrid.

The EMBO journal ·Vol. 9 ·No. 7 ·1990-07-00 ·Pages 2341-8

Skarstad K, Baker TA, Kornberg A

Abstract

The Escherichia coli initiator protein, dnaA complexed with ATP, binds to the replication origin (oriC) and melts the duplex in the AT-rich part of oriC. Opening of the duplex requires a minimum temperature and superhelix density. A low level of HU protein (5 molecules/oriC) is stimulatory, probably by facilitating DNA bending. High levels of HU protein (136 molecules/oriC) restrain the free superhelicity. Under adverse conditions for oriC melting (i.e. absence of HU, high levels of HU, low temperature or low superhelicity), transcription can activate initiation of replication by generating an R-loop located even a large distance from oriC. The strand-separation stage at oriC prior to the entry of the dnaB helicase was specifically stimulated. Activation by the R-loop was blocked when a stretch of 14 GC base pairs was inserted between the loop and oriC. Thus, activation of oriC by the distantly located R-loop appears to require propagation of DNA melting through the intervening sequence.

MeSH Terms
Bacterial Proteins/metabolism Base Sequence Chromosomes, Bacterial DNA Replication DNA, Bacterial/genetics,metabolism DNA-Binding Proteins/metabolism Escherichia coli/genetics,metabolism Kinetics Molecular Sequence Data Nucleic Acid Conformation Nucleic Acid Denaturation Nucleic Acid Hybridization Plasmids RNA, Bacterial/genetics,metabolism Restriction Mapping
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins RNA, Bacterial histone-like protein HU, bacteria
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Skarstad K
Department of Biochemistry, Stanford University School of Medicine, CA 94305-5307.
Baker T A
Kornberg A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-07-00
Pages
2341-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC551962
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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