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PMID: 2181438 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Escherichia coli Tus protein acts to arrest the progression of DNA replication forks in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 7 ·1990-04-00 ·Pages 2481-5

Hill TM, Marians KJ

Abstract

A polar DNA replication barrier is formed when the DNA-binding protein Tus forms a complex with any of the four 23-base-pair terminator (ter) sites found in the terminus region of the Escherichia coli chromosome. We have used a plasmid DNA replication system reconstituted with purified proteins in vitro to investigate the interaction of the Tus protein with the replication fork. Purified Tus protein alone is necessary and sufficient to arrest DNA replication on ColE1-type plasmid templates containing ter sites. Tus protein-catalyzed termination depends upon the orientation of the ter site in the plasmid DNA. Nucleotide resolution mapping of the terminated nascent DNA shows that leading-strand DNA synthesis arrests at the point of contact with the Tus protein, while the final lagging-strand primer sites are 50-70 nucleotides upstream. In addition, the distribution of leading-strand arrest sites changes when the composition of the proteins on the lagging-strand side of the replication fork is altered.

MeSH Terms
Bacterial Proteins/isolation & purification,metabolism Base Sequence DNA Replication DNA, Bacterial/biosynthesis,genetics Escherichia coli/genetics,metabolism Escherichia coli Proteins Kinetics Molecular Sequence Data Oligonucleotide Probes Plasmids Terminator Regions, Genetic
Chemicals
Bacterial Proteins DNA, Bacterial Escherichia coli Proteins Oligonucleotide Probes tus protein, Bacteria tus protein, E coli
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hill T M
Department of Biosciences and Biotechnology, Drexel University, Philadelphia, PA 19104.
Marians K J
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25 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.87.7.2481
Published
1990-04-00
Pages
2481-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC53713
Subset
IM
Grants
NIGMS NIH HHS · GM34558 · United States
NIGMS NIH HHS · GM43193 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.7.2481 PMC Full Text View journal details
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