DNA binding of PriA protein requires cooperation of the N-terminal D-loop/arrested-fork binding and C-terminal helicase domains

Taku Tanaka, Toshimi Mizukoshi, Chika Taniyama, Daisuke Kohda, Ken Ichi Arai, Hisao Masai

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

PriA protein is essential for RecA-dependent DNA replication induced by stalled replication forks in Escherichia coli. PriA is a DEXH-type DNA helicase, ATPase activity of which depends on its binding to structured DNA including a D-loop-like structure. Here, we show that the N-terminal 181-amino acid polypeptide can form a complex with D-loop in gel shift assays and have identified a unique motif present in the N-terminal segment of PriA that plays a role in its DNA binding. We have also identified residues in the C terminus proximal helicase domain essential for D-loop binding. PriA proteins mutated in this domain do not bind to D-loop, despite the presence of the N-terminal DNA-binding motif. Those mutants that cannot bind to D-loop in vitro do not support a recombination-dependent mode of DNA replication in vivo, indicating that binding to a D-loop-like structure is essential for the ability of PriA to initiate DNA replication and repair from stalled replication forks. We propose that binding of the PriA protein to stalled replication forks requires proper configuration of the N-terminal fork-recognition and C-terminal helicase domains and that the latter may stabilize binding and increase binding specificity.

Original languageEnglish
Pages (from-to)38062-38071
Number of pages10
JournalJournal of Biological Chemistry
Volume277
Issue number41
DOIs
Publication statusPublished - Oct 11 2002
Externally publishedYes

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DNA-Binding Proteins
DNA Replication
DNA
DNA Helicases
Nucleotide Motifs
Proteins
DNA Repair
Genetic Recombination
Adenosine Triphosphatases
Carrier Proteins
Gels
Escherichia coli
Amino Acids
Peptides
Assays
Repair

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

DNA binding of PriA protein requires cooperation of the N-terminal D-loop/arrested-fork binding and C-terminal helicase domains. / Tanaka, Taku; Mizukoshi, Toshimi; Taniyama, Chika; Kohda, Daisuke; Arai, Ken Ichi; Masai, Hisao.

In: Journal of Biological Chemistry, Vol. 277, No. 41, 11.10.2002, p. 38062-38071.

Research output: Contribution to journalArticle

Tanaka, Taku ; Mizukoshi, Toshimi ; Taniyama, Chika ; Kohda, Daisuke ; Arai, Ken Ichi ; Masai, Hisao. / DNA binding of PriA protein requires cooperation of the N-terminal D-loop/arrested-fork binding and C-terminal helicase domains. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 41. pp. 38062-38071.
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