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PMID: 2961561 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.

Evidence that the normal route of replication-allowed Red-mediated recombination involves double-chain ends.

The EMBO journal ·Vol. 6 ·No. 10 ·1987-10-00 ·Pages 3171-6

Thaler DS, Stahl MM, Stahl FW

Abstract

Recombination mediated by the Red pathway of bacteriophage lambda is focused towards sites of double-chain cuts. Double-chain ends created either by type II restriction enzymes acting at unmodified recognition sites or by lambda's packaging enzyme, terminase, acting at cos are utilized in a manner similar to the double-chain break repair pathway of recombination in yeast. When lambda is allowed to recombine during replicative growth, spontaneous recombination is approximately evenly distributed along the chromosome. It has been proposed that replication-allowed recombination also is initiated by double-chain ends. In order to test this hypothesis we ask if the in vivo expression of the Mu gam protein is inhibitory to Red recombination. Mu gam has been shown in vitro to bind to linearized duplex DNA and to shield bound DNA from exonucleases. The expression of Mu gam is found to be inhibitory to Red recombination whether replication is blocked or allowed. As a control we ask if Mu gam inhibits Int-mediated recombination. It has been well documented that the Int pathway of recombination does not involve any double-chain breaks and, consistent with this, the Int pathway is not inhibited by Mu gam. We suggest that the in vivo expression of Mu gam or other similar activities may be a generally useful way to determine if those processes that respond to an artificially introduced double-chain cut normally involve double-chain ends.

MeSH Terms
Bacteriophage lambda/genetics Crosses, Genetic DNA Replication DNA, Viral/genetics Escherichia coli/genetics Nucleic Acid Heteroduplexes/genetics Plasmids Recombination, Genetic Temperature
Chemicals
DNA, Viral Nucleic Acid Heteroduplexes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Thaler D S
Institute of Molecular Biology, University of Oregon, Eugene 97403.
Stahl M M
Stahl F W
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1987-10-00
Pages
3171-6
Language
English
Region
England
NLM ID
8208664
PMCID
PMC553759
Subset
IM
Grants
NIGMS NIH HHS · GM 07759 · United States
NIGMS NIH HHS · GM 33677 · United States
External Links
PubMed source PMC Full Text View journal details
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