Identification of a potential membrane-targeting region of the replication initiator protein (TrfA) of broad-host-range plasmid RK2.

Abstract:

:Plasmid RK2 codes for two species of the replication initiator protein TrfA (33 and 44 kDa). Both polypeptides are strongly associated with membrane fractions of Escherichia coli host cells (W. Firshein and P. Kim, Mol. Microbiol. 23, 1-10, 1997). We investigated the role of a 12-amino-acid hydrophobic region (HR) in the membrane association of TrfA. Epitope-tagged polypeptide fragments of TrfA that contained HR were expressed and found to be associated with membrane fractions. Site-directed mutagenesis of trfA revealed that changes of specific amino acids in HR can affect both TrfA association with the membrane and its ability to support replication of an RK2 oriV plasmid in vivo. These results are consistent with the hypothesis that membrane association of TrfA is functionally relevant and that the HR region of TrfA is involved in membrane association and DNA replication in vivo.

journal_name

Plasmid

journal_title

Plasmid

authors

Kim PD,Rosche TM,Firshein W

doi

10.1006/plas.2000.1467

keywords:

subject

Has Abstract

pub_date

2000-05-01 00:00:00

pages

214-22

issue

3

eissn

0147-619X

issn

1095-9890

pii

S0147-619X(00)91467-7

journal_volume

43

pub_type

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