Uncoupling of the DNA topoisomerase and replication activities of an initiator protein.

Abstract:

:The replication initiator proteins encoded by the pT181 and related plasmids have sequence-specific DNA binding and topoisomerase activities. These proteins create a site-specific nick in one strand of the DNA at the origin of replication that serves as a primer for the initiation of replication. To define the regions of the pT181-encoded initiator protein, RepC, that are involved in its DNA binding, topoisomerase, and replication activities, we have carried out site-directed mutagenesis of the repC gene. Analysis of mutant RepC proteins in vitro and in vivo has identified the amino acids that are critical for its various biochemical activities. The DNA binding domain of RepC was found to be located near its C-terminal region and was different from the domain involved in its sequence-specific topoisomerase activity. These studies also showed that the DNA topoisomerase activity of the initiator protein can be uncoupled from its tight noncovalent DNA binding and replication activities.

authors

Dempsey LA,Birch P,Khan SA

doi

10.1073/pnas.89.7.3083

keywords:

subject

Has Abstract

pub_date

1992-04-01 00:00:00

pages

3083-7

issue

7

eissn

0027-8424

issn

1091-6490

journal_volume

89

pub_type

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