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PMID: 2168553 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro assembly of relaxosomes at the transfer origin of plasmid RP4.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 17 ·1990-09-00 ·Pages 6555-9

Pansegrau W, Balzer D, Kruft V, Lurz R, Lanka E

Abstract

During initiation of conjugative transfer of DNA containing the transfer origin (oriT) of the promiscuous plasmid RP4, the proteins TraI, TraJ, and TraH interact and assemble a specialized nucleoprotein complex (the relaxosome) at oriT. The structure can be visualized on electron micrographs. Site- and strand-specific nicking at the transfer origin in vitro is dependent on the proteins TraI and TraJ and on Mg2+ ions. Substrate specificity is directed exclusively towards the cognate transfer origin: the RP4-specified TraJ protein cannot recognize the closely related oriT of plasmid R751. After nicking, TraI protein remains attached to the 5'-terminal 2'-deoxycytidyl residue at the nic site [Pansegrau, W., Ziegelin, G. & Lanka, E. (1990) J. Biol. Chem. 265, 10637-10644]. Nicking and relaxosome formation require supercoiled DNA. Thus, a complicated structure involving multiple plasmid-specified proteins and a defined region of DNA must be formed at the transfer origin to prepare the plasmid for generating the single strand to be transferred.

MeSH Terms

Amino Acid Sequence Bacterial Outer Membrane Proteins/genetics,metabolism Bacterial Proteins Base Sequence Cloning, Molecular Conjugation, Genetic DNA Helicases/genetics,metabolism DNA, Superhelical/genetics,ultrastructure Escherichia coli/genetics Escherichia coli Proteins Molecular Sequence Data Nuclear Proteins/genetics,metabolism Nucleoproteins/metabolism,ultrastructure R Factors Restriction Mapping

Chemicals

Bacterial Outer Membrane Proteins Bacterial Proteins DNA, Superhelical Escherichia coli Proteins Nuclear Proteins Nucleoproteins TraH protein, Bacteria TraJ protein, E coli TraJ protein, bacteria TraI protein, E coli DNA Helicases

Authors & Affiliations

5 authors, click to expand affiliations / ORCID

Pansegrau W

Max-Planck-Institut für Molekulare Genetik, Abteilung Schuster, Berlin, Federal Republic of Germany.

Balzer D

Kruft V

Lurz R

Lanka E

References (12)

12 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.87.17.6555

Published

1990-09-00

Pages

6555-9

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC54575

Subset

External Links

PubMed source DOI: 10.1073/pnas.87.17.6555 PMC Full Text View journal details

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In vitro assembly of relaxosomes at the transfer origin of plasmid RP4.

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