The Bacillus subtilis chromatin-associated protein Hbsu is involved in DNA repair and recombination

被引:37
作者
Fernandez, S [1 ]
Rojo, F [1 ]
Alonso, JC [1 ]
机构
[1] UNIV AUTONOMA MADRID,CSIC,CTR NACL BIOTECNOL,E-28049 MADRID,SPAIN
关键词
D O I
10.1046/j.1365-2958.1997.3061670.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Bacillus subtilis hbs gene encodes an essential chromatin-associated protein termed Hbsu. Hbsu, the counterpart of the Escherichia coli HU protein, binds DNA in a non-specific way but has a clear preference for bent, kinked or altered DNA sequences. To investigate the role of Hbsu in DNA repair and DNA recombination we have constructed a series of site-directed mutants in the hbs gene and used these mutant genes to substitute the wild-type chromosomal hbs gene. The hbs47 mutation, which codes for a mutant protein in which residue Phe-47 has been replaced by Trp, does not cause any discernible phenotype. Additional substitution of residue Arg-55 by Ala (hbs4755 mutation) rendered cells deficient in DNA repair, homologous recombination and beta protein-mediated site-specific recombination. We have also tested the effect on DNA repair of the hbs4755 mutation in combination with mutations in different functions of homologous DNA recombination (recA, recF, recG, recH and addAB). The hbs4755 mutation did not modify the sensitivity of recH and addAB cells to the DNA-damaging agents methylmethane sulphonate (MMS) or 4-nitroquinoline-1-oxide (4NQO), and it only marginally affected recF and recG cells. The hbs4755 mutation blocked intermolecular recombination in recH cells and markedly reduced it (20- to 50-fold) in recF and recG cells, but had no effect on addAB cells. Taken together, these data indicate that the Hbsu protein is required for DNA repair and for homologous DNA recombination.
引用
收藏
页码:1169 / 1179
页数:11
相关论文
共 44 条
[1]   THE ROLE OF THE CHROMATIN-ASSOCIATED PROTEIN HBSU IN BETA-MEDIATED DNA RECOMBINATION IS TO FACILITATE THE JOINING OF DISTANT RECOMBINATION SITES [J].
ALONSO, JC ;
GUTIERREZ, C ;
ROJO, F .
MOLECULAR MICROBIOLOGY, 1995, 18 (03) :471-478
[2]   THE BACILLUS-SUBTILIS HISTONE-LIKE PROTEIN HBSU IS REQUIRED FOR DNA RESOLUTION AND DNA INVERSION MEDIATED BY THE BETA-RECOMBINASE OF PLASMID PSM19035 [J].
ALONSO, JC ;
WEISE, F ;
ROJO, F .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (07) :2938-2945
[3]   GENETIC-RECOMBINATION IN BACILLUS-SUBTILIS 168 - EFFECT OF RECN, RECF, RECH AND ADDAB MUTATIONS ON DNA-REPAIR AND RECOMBINATION [J].
ALONSO, JC ;
STIEGE, AC ;
LUDER, G .
MOLECULAR & GENERAL GENETICS, 1993, 239 (1-2) :129-136
[4]  
Alonso JC, 1996, FEMS MICROBIOL LETT, V142, P1
[5]   CHARACTERIZATION OF RECOMBINATION-DEFICIENT MUTANTS OF BACILLUS-SUBTILIS [J].
ALONSO, JC ;
TAILOR, RH ;
LUDER, G .
JOURNAL OF BACTERIOLOGY, 1988, 170 (07) :3001-3007
[6]  
ALONSO JC, 1996, MICROBIOLOGIA GENETI, P229
[7]   RecR is a zinc metalloprotein from Bacillus subtilis 168 [J].
Ayora, S ;
Stiege, AC ;
Alonso, JC .
MOLECULAR MICROBIOLOGY, 1997, 23 (04) :639-647
[8]   PROKARYOTIC HU AND EUKARYOTIC HMG1 - A KINKED RELATIONSHIP [J].
BIANCHI, ME .
MOLECULAR MICROBIOLOGY, 1994, 14 (01) :1-5
[9]   DNA-BINDING PARAMETERS OF THE HU PROTEIN OF ESCHERICHIA-COLI TO CRUCIFORM DNA [J].
BONNEFOY, E ;
TAKAHASHI, M ;
YANIV, JR .
JOURNAL OF MOLECULAR BIOLOGY, 1994, 242 (02) :116-129
[10]   INCREASED SENSITIVITY TO GAMMA-IRRADIATION IN BACTERIA LACKING PROTEIN HU [J].
BOUBRIK, F ;
ROUVIEREYANIV, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (09) :3958-3962