Stability of linear DNA in recA mutant Escherichia coli cells reflects ongoing chromosomal DNA degradation

被引:49
作者
Kuzminov, A
Stahl, FW
机构
[1] Institute of Molecular Biology, University of Oregon, Eugene
关键词
D O I
10.1128/jb.179.3.880-888.1997
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
To study the fate of linear DNA in Escherichia coli cells, we linearized plasmid DNA at a specific site in vivo and monitored its behavior in recA mutant cells deficient in recombinational repair. Earlier, we had found that in wild-type (WT) cells linearized DNA is degraded to completion by RecBCD nuclease. We had also found that in WT cells chi sites on linear DNA inhibit RecBCD degradation by turning off its nucleolytic activities, Now we report that chi sites do not work in the absence of the RecA protein, suggesting that RecA is required in vivo to turn off the degradative activities of the RecBCD enzyme. We also report that the degradation of linearized plasmid DNA, even devoid of chi sites, is never complete in recA cells, Investigation of this linear DNA stability indicates that a fraction of recA cells are recBC phenocopies due to ongoing chromosomal DNA degradation, which titrates RecBCD nuclease, A possible role for RecBCD-promoted DNA degradation in controlling chromosomal DNA replication in E. coli is discussed.
引用
收藏
页码:880 / 888
页数:9
相关论文
共 56 条
  • [1] RECD - THE GENE FOR AN ESSENTIAL 3RD SUBUNIT OF EXONUCLEASE-V
    AMUNDSEN, SK
    TAYLOR, AF
    CHAUDHURY, AM
    SMITH, GR
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (15) : 5558 - 5562
  • [2] PEDIGREES OF SOME MUTANT STRAINS OF ESCHERICHIA-COLI K-12
    BACHMANN, BJ
    [J]. BACTERIOLOGICAL REVIEWS, 1972, 36 (04) : 525 - 557
  • [3] BACTERIOPHAGE-LAMBDA DNA - THE BEGINNING OF THE END
    BECKER, A
    MURIALDO, H
    [J]. JOURNAL OF BACTERIOLOGY, 1990, 172 (06) : 2819 - 2824
  • [4] BIRNBOIM HC, 1983, METHOD ENZYMOL, V100, P243
  • [5] INTERACTION OF RECBCD ENZYME WITH DNA DAMAGED BY GAMMA-RADIATION
    BRCICKOSTIC, K
    SALAJSMIC, E
    MARSIC, N
    KAJIC, S
    STOJILJKOVIC, I
    TRGOVCEVIC, Z
    [J]. MOLECULAR AND GENERAL GENETICS, 1991, 228 (1-2): : 136 - 142
  • [6] DNA CONTENT, SYNTHESIS AND INTEGRITY IN DIVIDING AND NON-DIVIDING CELLS OF REC-STRAINS OF ESCHERICHIA-COLI-K12
    CAPALDO, FN
    BARBOUR, SD
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1975, 91 (01) : 53 - 66
  • [7] ANALYSIS OF GROWTH OF RECOMBINATION-DEFICIENT STRAINS OF ESCHERICHIA-COLI-K-12
    CAPALDO, FN
    RAMSEY, G
    BARBOUR, SD
    [J]. JOURNAL OF BACTERIOLOGY, 1974, 118 (01) : 242 - 249
  • [8] Carlson Karin, 1994, P421
  • [9] THE PMTL NIC-CLONING VECTORS .1. IMPROVED PUC POLYLINKER REGIONS TO FACILITATE THE USE OF SONICATED DNA FOR NUCLEOTIDE SEQUENCING
    CHAMBERS, SP
    PRIOR, SE
    BARSTOW, DA
    MINTON, NP
    [J]. GENE, 1988, 68 (01) : 139 - 149
  • [10] A NEW CLASS OF ESCHERICHIA-COLI RECBC MUTANTS - IMPLICATIONS FOR THE ROLE OF RECBC ENZYME IN HOMOLOGOUS RECOMBINATION
    CHAUDHURY, AM
    SMITH, GR
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (24): : 7850 - 7854