General mechanism for RecA protein binding to duplex DNA

Benjamin Franklin Pugh, Michael M. Cox

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

RecA protein binding to duplex DNA occurs by a multi-step process. The tau analysis, originally developed to examine the binding of RNA polymerase to promoter DNA, is adapted here to study two kinetically distinguishable reaction segments of RecA-double stranded (ds) DNA complex formation in greater detail. One, which is probably a rapid preequilibrium in which RecA protein binds weakly to native dsDNA, is found to have the following properties: (1) a sensitivity to pH, involving a net release of approximately one proton; (2) a sensitivity to salts; (3) little or no dependence on temperature; (4) little or no dependence on DNA length. The second reaction segment, the rate-limiting nucleation of nucleoprotein filament formation accompanied by partial DNA unwinding, is found to have the following properties: (1) a sensitivity to pH, involving a net uptake of approximately three protons; (2) a sensitivity to salts; (3) a relatively large dependence on temperature, with an Arrhenius activation energy of 39 kcal mol-1; (4) a sensitivity to DNA topology; (5) a dependence on DNA length. These results contribute to a general mechanism for RecA protein binding to duplex DNA, which can provide a rationale for the apparent preferential binding to altered DNA structures such as pyrimidine dimers and Z-DNA.

Original languageEnglish (US)
Pages (from-to)479-493
Number of pages15
JournalJournal of Molecular Biology
Volume203
Issue number2
DOIs
StatePublished - Sep 20 1988

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Rec A Recombinases
Protein Binding
DNA
Protons
Salts
Z-Form DNA
Pyrimidine Dimers
Temperature
Nucleoproteins
DNA-Directed RNA Polymerases

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

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General mechanism for RecA protein binding to duplex DNA. / Pugh, Benjamin Franklin; Cox, Michael M.

In: Journal of Molecular Biology, Vol. 203, No. 2, 20.09.1988, p. 479-493.

Research output: Contribution to journalArticle

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