The directionality of RuvAB-mediated branch migration: In vitro studies with three-armed junctions

被引:19
作者
Hiom, K [1 ]
Tsaneva, IR [1 ]
West, SC [1 ]
机构
[1] IMPERIAL CANC RES FUND, CLARE HALL LABS, S MIMMS EN6 3LD, HERTS, ENGLAND
关键词
D O I
10.1046/j.1365-2443.1996.d01-253.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Background: The Escherichia coli RuvA and RuvB proteins promote the branch migration of 4-way (Holliday) junctions during genetic recombination. The active complex is a tripartite structure in which RuvA protein is bound to the crossover and is sandwiched between two hexameric rings of RuvB. Branch migration requires ATP hydrolysis and occurs as the DNA passes through each RuvB ring. Results: In this work, we have investigated the mechanism by which RuvAB catalyses the branch migration of a three-armed (Y) junction. Using synthetic DNA structures, we observed the formation of DNA products, a partial duplex DNA molecule and a single-stranded oligonucleotide, indicative of a branch migration reaction that occurred with unique polarity. Analysis of the RuvAB-junction complex by DNase footprinting showed that RuvA bound asymmetrically to the junction and targeted a single hexameric RuvB ring to one arm of DNA. Conclusion: Branch migration of a three-armed junction occurs in a unidirectional manner that is determined by the assembly of a single RuvB ring onto one arm of the DNA. The asymmetry of the complex and observed directionality of branch migration indicate that strand passage occurs as the DNA is pulled into the RuvB ring structure, a reaction likely to be coupled with DNA unwinding.
引用
收藏
页码:443 / 451
页数:9
相关论文
共 27 条
  • [11] THE ESCHERICHIA-COLI RUVAB PROTEINS BRANCH MIGRATE HOLLIDAY JUNCTIONS THROUGH HETEROLOGOUS DNA-SEQUENCES IN A REACTION FACILITATED BY SSB
    PARSONS, CA
    STASIAK, A
    WEST, SC
    [J]. EMBO JOURNAL, 1995, 14 (22) : 5736 - 5744
  • [12] SPECIFICITY OF BINDING TO 4-WAY JUNCTIONS IN DNA BY BACTERIOPHAGE-T7 ENDONUCLEASE-I
    PARSONS, CA
    WEST, SC
    [J]. NUCLEIC ACIDS RESEARCH, 1990, 18 (15) : 4377 - 4384
  • [13] PARSONS CA, 1990, J BIOL CHEM, V265, P9285
  • [14] STRUCTURE OF A MULTISUBUNIT COMPLEX THAT PROMOTES DNA BRANCH MIGRATION
    PARSONS, CA
    STASIAK, A
    BENNETT, RJ
    WEST, SC
    [J]. NATURE, 1995, 374 (6520) : 375 - 378
  • [15] FORMATION OF A RUVAB-HOLLIDAY JUNCTION COMPLEX IN-VITRO
    PARSONS, CA
    WEST, SC
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1993, 232 (02) : 397 - 405
  • [16] INTERACTION OF ESCHERICHIA-COLI RUVA AND RUVB PROTEINS WITH SYNTHETIC HOLLIDAY JUNCTIONS
    PARSONS, CA
    TSANEVA, I
    LLOYD, RG
    WEST, SC
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (12) : 5452 - 5456
  • [17] Sambrook E, 1989, MOL CLONING LAB MANU
  • [18] MOLECULAR MECHANISMS OF HOLLIDAY JUNCTION PROCESSING IN ESCHERICHIA-COLI
    SHINAGAWA, H
    IWASAKI, H
    [J]. ADVANCES IN BIOPHYSICS, VOL 31, 1995, 1995, 31 : 49 - 65
  • [19] THE ESCHERICHIA-COLI RUVB BRANCH MIGRATION PROTEIN FORMS DOUBLE HEXAMERIC RINGS AROUND DNA
    STASIAK, A
    TSANEVA, IR
    WEST, SC
    BENSON, CJB
    YU, X
    EGELMAN, EH
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (16) : 7618 - 7622
  • [20] PURIFICATION AND PROPERTIES OF THE RUVA AND RUVB PROTEINS OF ESCHERICHIA-COLI
    TSANEVA, IR
    ILLING, G
    LLOYD, RG
    WEST, SC
    [J]. MOLECULAR & GENERAL GENETICS, 1992, 235 (01): : 1 - 10