Homologous pairing in vitro stimulated by the recombination Hotspot, Chi

Dan A. Dixon, Stephen C. Kowalczykowski

Research output: Contribution to journalArticle

115 Citations (Scopus)

Abstract

Genetic recombination in Escherichia coli is stimulated at DNA sequences known as Chi sites, 5′-GCT-GGTGG-3′. We describe the in vitro formation of homologously paired joint molecules that is dependent upon this recombination hotspot. Chi-dependent joint molecule formation requires RecA, RecBCD, and SSB proteins and a Chi site in the donor linear dsDNA. The donor dsDNA is unwound by RecBCD enzyme, and the invasive strand is generated by nicking at Chi. This Chi-dependent invading strand must contain homology to the recipient supercoiled DNA substrate at its newly formed 3′ end for efficient joint molecule formation. Action at Chi generates invasive ssDNA from the 5′ but not the 3′ side of Chi, suggesting that the nuclease activity of RecBCD enzyme is attenuated upon encountering a Chi site. These results support the view that RecBCD enzyme action can precede RecA protein action and reconcile the seemingly opposing degradative and recombination functions of RecBCD enzyme.

Original languageEnglish (US)
Pages (from-to)361-371
Number of pages11
JournalCell
Volume66
Issue number2
DOIs
StatePublished - Jul 26 1991
Externally publishedYes

Fingerprint

Exodeoxyribonuclease V
Genetic Recombination
Joints
Molecules
Rec A Recombinases
Superhelical DNA
DNA sequences
Escherichia coli
In Vitro Techniques
Substrates
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Homologous pairing in vitro stimulated by the recombination Hotspot, Chi. / Dixon, Dan A.; Kowalczykowski, Stephen C.

In: Cell, Vol. 66, No. 2, 26.07.1991, p. 361-371.

Research output: Contribution to journalArticle

Dixon, Dan A. ; Kowalczykowski, Stephen C. / Homologous pairing in vitro stimulated by the recombination Hotspot, Chi. In: Cell. 1991 ; Vol. 66, No. 2. pp. 361-371.
@article{e780e102dd5c4fe6924d168b0f7bcdff,
title = "Homologous pairing in vitro stimulated by the recombination Hotspot, Chi",
abstract = "Genetic recombination in Escherichia coli is stimulated at DNA sequences known as Chi sites, 5′-GCT-GGTGG-3′. We describe the in vitro formation of homologously paired joint molecules that is dependent upon this recombination hotspot. Chi-dependent joint molecule formation requires RecA, RecBCD, and SSB proteins and a Chi site in the donor linear dsDNA. The donor dsDNA is unwound by RecBCD enzyme, and the invasive strand is generated by nicking at Chi. This Chi-dependent invading strand must contain homology to the recipient supercoiled DNA substrate at its newly formed 3′ end for efficient joint molecule formation. Action at Chi generates invasive ssDNA from the 5′ but not the 3′ side of Chi, suggesting that the nuclease activity of RecBCD enzyme is attenuated upon encountering a Chi site. These results support the view that RecBCD enzyme action can precede RecA protein action and reconcile the seemingly opposing degradative and recombination functions of RecBCD enzyme.",
author = "Dixon, {Dan A.} and Kowalczykowski, {Stephen C.}",
year = "1991",
month = "7",
day = "26",
doi = "10.1016/0092-8674(91)90625-9",
language = "English (US)",
volume = "66",
pages = "361--371",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "2",

}

TY - JOUR

T1 - Homologous pairing in vitro stimulated by the recombination Hotspot, Chi

AU - Dixon, Dan A.

AU - Kowalczykowski, Stephen C.

PY - 1991/7/26

Y1 - 1991/7/26

N2 - Genetic recombination in Escherichia coli is stimulated at DNA sequences known as Chi sites, 5′-GCT-GGTGG-3′. We describe the in vitro formation of homologously paired joint molecules that is dependent upon this recombination hotspot. Chi-dependent joint molecule formation requires RecA, RecBCD, and SSB proteins and a Chi site in the donor linear dsDNA. The donor dsDNA is unwound by RecBCD enzyme, and the invasive strand is generated by nicking at Chi. This Chi-dependent invading strand must contain homology to the recipient supercoiled DNA substrate at its newly formed 3′ end for efficient joint molecule formation. Action at Chi generates invasive ssDNA from the 5′ but not the 3′ side of Chi, suggesting that the nuclease activity of RecBCD enzyme is attenuated upon encountering a Chi site. These results support the view that RecBCD enzyme action can precede RecA protein action and reconcile the seemingly opposing degradative and recombination functions of RecBCD enzyme.

AB - Genetic recombination in Escherichia coli is stimulated at DNA sequences known as Chi sites, 5′-GCT-GGTGG-3′. We describe the in vitro formation of homologously paired joint molecules that is dependent upon this recombination hotspot. Chi-dependent joint molecule formation requires RecA, RecBCD, and SSB proteins and a Chi site in the donor linear dsDNA. The donor dsDNA is unwound by RecBCD enzyme, and the invasive strand is generated by nicking at Chi. This Chi-dependent invading strand must contain homology to the recipient supercoiled DNA substrate at its newly formed 3′ end for efficient joint molecule formation. Action at Chi generates invasive ssDNA from the 5′ but not the 3′ side of Chi, suggesting that the nuclease activity of RecBCD enzyme is attenuated upon encountering a Chi site. These results support the view that RecBCD enzyme action can precede RecA protein action and reconcile the seemingly opposing degradative and recombination functions of RecBCD enzyme.

UR - http://www.scopus.com/inward/record.url?scp=0025902330&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025902330&partnerID=8YFLogxK

U2 - 10.1016/0092-8674(91)90625-9

DO - 10.1016/0092-8674(91)90625-9

M3 - Article

C2 - 1855256

AN - SCOPUS:0025902330

VL - 66

SP - 361

EP - 371

JO - Cell

JF - Cell

SN - 0092-8674

IS - 2

ER -