PcrA-mediated disruption of RecA nucleoprotein filaments-essential role of the ATPase activity of RecA

Matt V. Fagerburg; Grant D. Schauer; Karen R. Thickman; Piero R. Bianco; Saleem A. Khan; Sanford H. Leuba; Syam P. Anand

doi:10.1093/nar/gks641

PcrA-mediated disruption of RecA nucleoprotein filaments-essential role of the ATPase activity of RecA

Matt V. Fagerburg, Grant D. Schauer, Karen R. Thickman, Piero R. Bianco, Saleem A. Khan, Sanford H. Leuba, Syam P. Anand

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

The essential DNA helicase, PcrA, regulates recombination by displacing the recombinase RecA from the DNA. The nucleotide-bound state of RecA determines the stability of its nucleoprotein filaments. Using single-molecule fluorescence approaches, we demonstrate that RecA displacement by a translocating PcrA requires the ATPase activity of the recombinase. We also show that in a 'head-on collision' between a polymerizing RecA filament and a translocating PcrA, the RecA K72R ATPase mutant, but not wild-type RecA, arrests helicase translocation. Our findings demonstrate that translocation of PcrA is not sufficient to displace RecA from the DNA and assigns an essential role for the ATPase activity of RecA in helicase-mediated disruption of its filaments.

Original language	English (US)
Pages (from-to)	8416-8424
Number of pages	9
Journal	Nucleic acids research
Volume	40
Issue number	17
DOIs	https://doi.org/10.1093/nar/gks641
State	Published - Sep 2012
Externally published	Yes

ASJC Scopus subject areas

Genetics

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title = "PcrA-mediated disruption of RecA nucleoprotein filaments-essential role of the ATPase activity of RecA",

abstract = "The essential DNA helicase, PcrA, regulates recombination by displacing the recombinase RecA from the DNA. The nucleotide-bound state of RecA determines the stability of its nucleoprotein filaments. Using single-molecule fluorescence approaches, we demonstrate that RecA displacement by a translocating PcrA requires the ATPase activity of the recombinase. We also show that in a 'head-on collision' between a polymerizing RecA filament and a translocating PcrA, the RecA K72R ATPase mutant, but not wild-type RecA, arrests helicase translocation. Our findings demonstrate that translocation of PcrA is not sufficient to displace RecA from the DNA and assigns an essential role for the ATPase activity of RecA in helicase-mediated disruption of its filaments.",

author = "Fagerburg, {Matt V.} and Schauer, {Grant D.} and Thickman, {Karen R.} and Bianco, {Piero R.} and Khan, {Saleem A.} and Leuba, {Sanford H.} and Anand, {Syam P.}",

note = "Funding Information: National Institutes of Health (NIH) [R01GM31685 to S.A.K., R01GM077872 to S.H.L]; National Institute of General Medical Sciences fellowship [F32GM082132 to K.R.T.]. Funding for open access charge: NIH [R01GM31685 to S.A.K.; R01GM077872 to S.H.L.].",

year = "2012",

month = sep,

doi = "10.1093/nar/gks641",

language = "English (US)",

volume = "40",

pages = "8416--8424",

journal = "Nucleic acids research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "17",

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T1 - PcrA-mediated disruption of RecA nucleoprotein filaments-essential role of the ATPase activity of RecA

AU - Fagerburg, Matt V.

AU - Schauer, Grant D.

AU - Thickman, Karen R.

AU - Bianco, Piero R.

AU - Khan, Saleem A.

AU - Leuba, Sanford H.

AU - Anand, Syam P.

N1 - Funding Information: National Institutes of Health (NIH) [R01GM31685 to S.A.K., R01GM077872 to S.H.L]; National Institute of General Medical Sciences fellowship [F32GM082132 to K.R.T.]. Funding for open access charge: NIH [R01GM31685 to S.A.K.; R01GM077872 to S.H.L.].

PY - 2012/9

Y1 - 2012/9

N2 - The essential DNA helicase, PcrA, regulates recombination by displacing the recombinase RecA from the DNA. The nucleotide-bound state of RecA determines the stability of its nucleoprotein filaments. Using single-molecule fluorescence approaches, we demonstrate that RecA displacement by a translocating PcrA requires the ATPase activity of the recombinase. We also show that in a 'head-on collision' between a polymerizing RecA filament and a translocating PcrA, the RecA K72R ATPase mutant, but not wild-type RecA, arrests helicase translocation. Our findings demonstrate that translocation of PcrA is not sufficient to displace RecA from the DNA and assigns an essential role for the ATPase activity of RecA in helicase-mediated disruption of its filaments.

AB - The essential DNA helicase, PcrA, regulates recombination by displacing the recombinase RecA from the DNA. The nucleotide-bound state of RecA determines the stability of its nucleoprotein filaments. Using single-molecule fluorescence approaches, we demonstrate that RecA displacement by a translocating PcrA requires the ATPase activity of the recombinase. We also show that in a 'head-on collision' between a polymerizing RecA filament and a translocating PcrA, the RecA K72R ATPase mutant, but not wild-type RecA, arrests helicase translocation. Our findings demonstrate that translocation of PcrA is not sufficient to displace RecA from the DNA and assigns an essential role for the ATPase activity of RecA in helicase-mediated disruption of its filaments.

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JO - Nucleic acids research

JF - Nucleic acids research

IS - 17

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PcrA-mediated disruption of RecA nucleoprotein filaments-essential role of the ATPase activity of RecA

Abstract

ASJC Scopus subject areas

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