Interaction of the RecA protein of Escherichia coli with single-stranded oligodeoxyribonucleotides

Piero R. Bianco; George M. Weinstock

doi:10.1093/nar/24.24.4933

Interaction of the RecA protein of Escherichia coli with single-stranded oligodeoxyribonucleotides

Piero R. Bianco, George M. Weinstock

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

32 Scopus citations

Abstract

The RecA protein of Escherichia coli performs a number of ATP-dependent, in vitro reactions and is a DNA-dependent ATPase. Small oligodeoxyribonucleotides were used as DNA cofactors in a kinetic analysis of the ATPase reaction. Polymers of deoxythymidilic acid as well as oligonucleotides of mixed base composition stimulated the RecA ATPase activity in a length-dependent fashion. Both the initial rate and the extent of the reaction were affected by chain length. Full activity was seen with chain lengths ≤ 30 nt. Partial activity was seen with chain lengths of 15-30 nt. The lower activity of shorter oligonucleotides was not simply due to a reduced affinity for DNA, since effects of chain length on K(m)(ATP) and the Hill coefficient for ATP hydrolysis were also observed. The results also suggested that single-stranded DNA secondary structure frequently affects the ATPase activity of RecA protein with oligodeoxyribonucleotides.

Original language	English (US)
Pages (from-to)	4933-4939
Number of pages	7
Journal	Nucleic acids research
Volume	24
Issue number	24
DOIs	https://doi.org/10.1093/nar/24.24.4933
State	Published - Dec 15 1996
Externally published	Yes

ASJC Scopus subject areas

Genetics

Access to Document

10.1093/nar/24.24.4933

Cite this

@article{0caa52376ff9467c8e46a32f1102138f,

title = "Interaction of the RecA protein of Escherichia coli with single-stranded oligodeoxyribonucleotides",

abstract = "The RecA protein of Escherichia coli performs a number of ATP-dependent, in vitro reactions and is a DNA-dependent ATPase. Small oligodeoxyribonucleotides were used as DNA cofactors in a kinetic analysis of the ATPase reaction. Polymers of deoxythymidilic acid as well as oligonucleotides of mixed base composition stimulated the RecA ATPase activity in a length-dependent fashion. Both the initial rate and the extent of the reaction were affected by chain length. Full activity was seen with chain lengths ≤ 30 nt. Partial activity was seen with chain lengths of 15-30 nt. The lower activity of shorter oligonucleotides was not simply due to a reduced affinity for DNA, since effects of chain length on K(m)(ATP) and the Hill coefficient for ATP hydrolysis were also observed. The results also suggested that single-stranded DNA secondary structure frequently affects the ATPase activity of RecA protein with oligodeoxyribonucleotides.",

author = "Bianco, {Piero R.} and Weinstock, {George M.}",

note = "Funding Information: This work was supported in part by PHS grant GM3524 to G.M.W. This work was submitted in partial fulfillment of the PhD degree by P.R.B.",

year = "1996",

month = dec,

day = "15",

doi = "10.1093/nar/24.24.4933",

language = "English (US)",

volume = "24",

pages = "4933--4939",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "24",

}

TY - JOUR

T1 - Interaction of the RecA protein of Escherichia coli with single-stranded oligodeoxyribonucleotides

AU - Bianco, Piero R.

AU - Weinstock, George M.

N1 - Funding Information: This work was supported in part by PHS grant GM3524 to G.M.W. This work was submitted in partial fulfillment of the PhD degree by P.R.B.

PY - 1996/12/15

Y1 - 1996/12/15

N2 - The RecA protein of Escherichia coli performs a number of ATP-dependent, in vitro reactions and is a DNA-dependent ATPase. Small oligodeoxyribonucleotides were used as DNA cofactors in a kinetic analysis of the ATPase reaction. Polymers of deoxythymidilic acid as well as oligonucleotides of mixed base composition stimulated the RecA ATPase activity in a length-dependent fashion. Both the initial rate and the extent of the reaction were affected by chain length. Full activity was seen with chain lengths ≤ 30 nt. Partial activity was seen with chain lengths of 15-30 nt. The lower activity of shorter oligonucleotides was not simply due to a reduced affinity for DNA, since effects of chain length on K(m)(ATP) and the Hill coefficient for ATP hydrolysis were also observed. The results also suggested that single-stranded DNA secondary structure frequently affects the ATPase activity of RecA protein with oligodeoxyribonucleotides.

AB - The RecA protein of Escherichia coli performs a number of ATP-dependent, in vitro reactions and is a DNA-dependent ATPase. Small oligodeoxyribonucleotides were used as DNA cofactors in a kinetic analysis of the ATPase reaction. Polymers of deoxythymidilic acid as well as oligonucleotides of mixed base composition stimulated the RecA ATPase activity in a length-dependent fashion. Both the initial rate and the extent of the reaction were affected by chain length. Full activity was seen with chain lengths ≤ 30 nt. Partial activity was seen with chain lengths of 15-30 nt. The lower activity of shorter oligonucleotides was not simply due to a reduced affinity for DNA, since effects of chain length on K(m)(ATP) and the Hill coefficient for ATP hydrolysis were also observed. The results also suggested that single-stranded DNA secondary structure frequently affects the ATPase activity of RecA protein with oligodeoxyribonucleotides.

UR - http://www.scopus.com/inward/record.url?scp=0030460712&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030460712&partnerID=8YFLogxK

U2 - 10.1093/nar/24.24.4933

DO - 10.1093/nar/24.24.4933

M3 - Article

C2 - 9016663

AN - SCOPUS:0030460712

VL - 24

SP - 4933

EP - 4939

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 24

ER -

Interaction of the RecA protein of Escherichia coli with single-stranded oligodeoxyribonucleotides

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this