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PMID: 2166943 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro maturation and encapsidation of the DNA of transposable Mu-like phage D108.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 16 ·1990-08-00 ·Pages 6092-6

Burns CM, Chan HL, DuBow MS

Abstract

Mu and D108 are related, temperate, transposable coliphages with unusual modes of DNA replication (transposition) and virion DNA maturation. These double-stranded DNA genomes replicate intrachromosomally and are matured and encapsidated linked to DNA sequences flanking the dispersed, integrated phage genomes. We have developed an in vitro system that employs crude lysates prepared from cells late in the Mu lytic cycle and that is proficient for both maturation and encapsidation of D108 DNA. Different forms of phage DNA were packaged at different efficiencies, with a circular pSC101::D108cts10 plasmid being most efficient, linearized plasmid less so, and mature virion DNA a poor substrate. The addition of purified D108 Ner protein to the reaction had no effect, whereas D108 repressor (c protein) inhibited the reaction. Escherichia coli integration host factor and D108 transposase proteins exerted an inhibitory effect on circular DNA substrates but had little effect on linear DNA packaging. This in vitro system, coupled with that developed for transposition, can now be used to biochemically dissect the protein and substrate requirements of these phages' DNA maturation pathway and the nature of the molecular switch between DNA transposition and encapsidation.

MeSH Terms
Bacteriophage mu/genetics Carrier Proteins/metabolism DNA Transposable Elements DNA, Viral/genetics,metabolism DNA-Binding Proteins/metabolism Escherichia coli/genetics Escherichia coli Proteins Integration Host Factors Kinetics Nucleotidyltransferases/metabolism Plasmids Protein Binding Repressor Proteins/metabolism Transposases
Chemicals
Carrier Proteins DNA Transposable Elements DNA, Viral DNA-Binding Proteins Escherichia coli Proteins Integration Host Factors Repressor Proteins integration host factor, E coli Nucleotidyltransferases Transposases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Burns C M
McGill University, Department of Microbiology and Immunology, Montreal, PQ, Canada.
Chan H L
DuBow M S
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37 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.16.6092
Published
1990-08-00
Pages
6092-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC54478
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.87.16.6092 PMC Full Text View journal details
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