SPECIFICITY DETERMINANTS IN THE ATTACHMENT SITES OF BACTERIOPHAGE-HK022 AND BACTERIOPHAGE-LAMBDA

被引:38
作者
NAGARAJA, R [1 ]
WEISBERG, RA [1 ]
机构
[1] NICHHD,MOLEC GENET LAB,MICROBIAL GENET SECT,BETHESDA,MD 20892
关键词
D O I
10.1128/jb.172.11.6540-6550.1990
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The Int proteins of bacteriophages HK022 and λ promote recombination between phage and bacterial attachment sites. Although the proteins and attachment sites of the two phages are similar, neither protein promotes efficient recombination between the pair of attachment sites used by the other phage. To analyze this difference in specificity, we constructed and characterized chimeric attachment sites in which segments of one site were replaced with corresponding segments of the other. Most such chimeras recombined with appropriate partner sites in vivo and in vitro, and their differential responses to the Int proteins of the two phages allowed us to locate determinants of the specificity difference in the bacterial attachment sites and a central segment of the phage attachment sites. The location of these determinants encompasses three of the four core-type binding sites for λ Int: C, B, and most importantly, B'. The regions corresponding to the C' core binding site and the arm-type binding sites of λ Int play no role in the specificity difference and, indeed, are well conserved in the two phages. We found, unexpectedly, that the effect of replacement of an Int-binding region on the recombinational potency of one chimeric site was reversed by a change of partner. This novel context effect suggests that postsynaptic interactions affect the specificity of recognition of attachment sites by Int.
引用
收藏
页码:6540 / 6550
页数:11
相关论文
共 40 条
[1]  
BAUER CE, 1989, GENETICS, V122, P727
[2]   EXTENT OF SEQUENCE HOMOLOGY REQUIRED FOR BACTERIOPHAGE-LAMBDA SITE-SPECIFIC RECOMBINATION [J].
BAUER, CE ;
GARDNER, JF ;
GUMPORT, RI .
JOURNAL OF MOLECULAR BIOLOGY, 1985, 181 (02) :187-197
[3]  
BIRNBOIM HC, 1979, NUCLEIC ACIDS RES, V7, P1513
[4]   THE MECHANISM OF PHAGE-LAMBDA SITE-SPECIFIC RECOMBINATION - SITE-SPECIFIC BREAKAGE OF DNA BY INT TOPOISOMERASE [J].
CRAIG, NL ;
NASH, HA .
CELL, 1983, 35 (03) :795-803
[5]   ESCHERICHIA-COLI INTEGRATION HOST FACTOR BINDS TO SPECIFIC SITES IN DNA [J].
CRAIG, NL ;
NASH, HA .
CELL, 1984, 39 (03) :707-716
[6]   MUTATIONS OF THE PHAGE-LAMBDA ATTACHMENT SITE ALTER THE DIRECTIONALITY OF RESOLUTION OF HOLLIDAY STRUCTURES [J].
DEMASSY, B ;
DORGAI, L ;
WEISBERG, RA .
EMBO JOURNAL, 1989, 8 (05) :1591-1599
[7]   ENZYMES AND SITES OF GENETIC-RECOMBINATION - STUDIES WITH GENE-3 ENDONUCLEASE OF PHAGE-T7 AND WITH SITE-AFFINITY MUTANTS OF PHAGE-LAMBDA [J].
DEMASSY, B ;
STUDIER, FW ;
DORGAI, L ;
APPELBAUM, E ;
WEISBERG, RA .
COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY, 1984, 49 :715-726
[8]  
DENTE L, 1985, DNA CLONING PRACTICA, V1, P101
[9]   AUTONOMOUS DNA-BINDING DOMAINS OF LAMBDA-INTEGRASE RECOGNIZE 2 DIFFERENT SEQUENCE FAMILIES [J].
DEVARGAS, LM ;
PARGELLIS, CA ;
HASAN, NM ;
BUSHMAN, EW ;
LANDY, A .
CELL, 1988, 54 (07) :923-929