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噬菌体HK022和λ附着位点的特异性决定因素

Specificity determinants in the attachment sites of bacteriophages HK022 and lambda.

作者信息

Nagaraja R, Weisberg R A

机构信息

Section on Microbial Genetics, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.

出版信息

J Bacteriol. 1990 Nov;172(11):6540-50. doi: 10.1128/jb.172.11.6540-6550.1990.

Abstract

The Int proteins of bacteriophages HK022 and lambda 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 lambda Int: C, B, and most importantly, B'. The regions corresponding to the C' core binding site and the arm-type binding sites of lambda 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.

摘要

噬菌体HK022和λ的整合蛋白促进噬菌体与细菌附着位点之间的重组。尽管这两种噬菌体的蛋白质和附着位点相似,但任何一种蛋白质都不能促进另一种噬菌体所使用的一对附着位点之间的高效重组。为了分析这种特异性差异,我们构建并表征了嵌合附着位点,其中一个位点的片段被另一个位点的相应片段所取代。大多数此类嵌合体在体内和体外都能与合适的伙伴位点重组,并且它们对两种噬菌体整合蛋白的不同反应使我们能够确定细菌附着位点和噬菌体附着位点中央片段中特异性差异的决定因素。这些决定因素的位置涵盖了λ整合蛋白的四个核心型结合位点中的三个:C、B,最重要的是B'。与λ整合蛋白的C'核心结合位点和臂型结合位点相对应的区域在特异性差异中不起作用,实际上,在这两种噬菌体中它们是高度保守的。我们意外地发现,一个嵌合位点的整合蛋白结合区域的替换对其重组效力的影响会因伙伴的改变而逆转。这种新的背景效应表明,突触后相互作用会影响整合蛋白对附着位点识别的特异性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9e25/526843/10fbd626e7cc/jbacter00165-0379-a.jpg

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