Isolation and Visualization of Active Presynaptic Filaments of RecA Protein and Single-stranded DNA

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1984 Nov 1

PMID 6594678

Citations 43

Authors

J Flory

S S Tsang

K Muniyappa

Affiliations

Soon will be listed here.

Abstract

In the presence of ATP, recA protein forms a presynaptic complex with single-stranded DNA that is an obligatory intermediate in homologous pairing. Presynaptic complexes of recA protein and circular single strands that are active in forming joint molecules can be isolated by gel filtration. These isolated active complexes are nucleoprotein filaments with the following characteristics: (i) a contour length that is at least 1.5 times that of the corresponding duplex DNA molecule, (ii) an ordered structure visualized by negative staining as a striated filament with a repeat distance of 9.0 nm and a width of 9.3 nm, (iii) approximately 8 molecules of recA protein and 20 nucleotide residues per striation. The widened spacing between bases in the nucleoprotein filament means that the initial matching of complementary sequences must involve intertwining of the filament and duplex DNA, unwinding of the latter, or some combination of both to equalize the spacing between nascent base pairs. These experiments support the concept that recA protein first forms a filament with single-stranded DNA, which in turn binds to duplex DNA to mediate both homologous pairing and subsequent strand exchange.

Citing Articles

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Prokaryotic DNA Crossroads: Holliday Junction Formation and Resolution.

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Homology recognition without double-stranded DNA-strand separation in D-loop formation by RecA.

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The intrinsic ability of double-stranded DNA to carry out D-loop and R-loop formation.

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Nonfilament-forming RecA dimer catalyzes homologous joint formation.

Shinohara T, Arai N, Iikura Y, Kasagi M, Masuda-Ozawa T, Yamaguchi Y Nucleic Acids Res. 2018; 46(20):10855-10869.

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