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PMID: 2826803 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The Nul subunit of bacteriophage lambda terminase binds to specific sites in cos DNA.

Journal of virology ·Vol. 62 ·No. 2 ·1988-02-00 ·Pages 387-92

Shinder G, Gold M

Abstract

The maturation and packaging of bacteriophage lambda DNA are under the control of the multifunctional viral terminase enzyme, which is composed of the protein products of Nu1 and A, the two most leftward genes of the phage chromosome. Terminase binds selectively to the cohesive end site (cos) of multimeric replicating lambda DNA and introduces staggered nicks to regenerate the 12-base single-stranded cohesive ends of the mature phage genome. The purified gpNu1 subunit of terminase forms specific complexes with cos lambda DNA. DNase I footprinting experiments showed that gpNu1 bound to three distinct regions near the extreme left end of the lambda chromosome. These regions coincided with two 16-base-pair sequences (CTGTCGTTTCCTTTCT) that were in inverted orientation, as well as a truncated version of this sequence. Bear et al. (J. Virol. 52:966-972,1984) isolated a mutant phage which contained a CG to TA transition at the 10th position of the rightmost 16-base-pair sequence, and this phage (termed lambda cos 154) exhibits a defect in DNA maturation when it replicates in Escherichia coli which is deficient in integration host factor. Footprinting experiments with cos 154 DNA showed that gpNu1 could not bind to the site which contained the mutation but could protect the other two sites. Since the DNA-packaging specificity of terminase resides in the gpNu1 subunit, these studies suggest that terminase uses these three sites as recognition sequences for specific binding to cos lambda.

MeSH Terms
Bacteriophage lambda/enzymology,genetics Base Sequence DNA Restriction Enzymes DNA, Viral/genetics,metabolism Deoxyribonuclease I/genetics Electrophoresis, Polyacrylamide Gel Endodeoxyribonucleases/genetics,metabolism Genes, Viral Molecular Sequence Data Mutation
Chemicals
DNA, Viral Endodeoxyribonucleases terminase DNA Restriction Enzymes Deoxyribonuclease I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shinder G
Department of Medical Genetics, University of Toronto, Ontario, Canada.
Gold M
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22 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
Published
1988-02-00
Pages
387-92
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC250547
Subset
IM
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