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通过改变DNA解旋酶II(uvrD)和IV(helD)来抑制大肠杆菌recJ核酸外切酶突变体。

Suppression of recJ exonuclease mutants of Escherichia coli by alterations in DNA helicases II (uvrD) and IV (helD).

作者信息

Lovett S T, Sutera V A

机构信息

Department of Biology, Brandeis University, Waltham, Massachusetts 02254-9110, USA.

出版信息

Genetics. 1995 May;140(1):27-45. doi: 10.1093/genetics/140.1.27.

Abstract

The recJ gene encodes a single-strand DNA-specific exonuclease involved in homologous recombination. We have isolated a pseudorevertant strain in which recJ mutant phenotypes were alleviated. Suppression of recJ was due to at least three mutations, two of which we have identified as alterations in DNA helicase genes. A recessive amber mutation, "uvrD517am," at codon 503 of the gene encoding helicase II was sufficient to suppress recJ partially. The uvrD517am mutation does not eliminate uvrD function because it affects UV survival only weakly; moreover, a uvrD insertion mutation could not replace uvrD517am as a suppressor. However, suppression may result from differential loss of uvrD function: mutation rate in a uvrD517am derivative was greatly elevated, equal to that in a uvrD insertion mutant. The second cosuppressor mutation is an allele of the helD gene, encoding DNA helicase IV, and could be replaced by insertion mutations in helD. The identity of the third cosuppressor "srjD" is not known. Strains carrying the three cosuppressor mutations exhibited hyperrecombinational phenotypes including elevated excision of repeated sequences. To explain recJ suppression, we propose that loss of antirecombinational helicase activity by the suppressor mutations stabilizes recombinational intermediates formed in the absence of recJ.

摘要

recJ基因编码一种参与同源重组的单链DNA特异性核酸外切酶。我们分离出了一个假回复突变菌株,其中recJ突变表型得到了缓解。recJ的抑制至少归因于三个突变,其中两个我们已确定为DNA解旋酶基因的改变。编码解旋酶II的基因第503位密码子处的隐性琥珀突变“uvrD517am”足以部分抑制recJ。uvrD517am突变不会消除uvrD功能,因为它对紫外线存活的影响很微弱;此外,uvrD插入突变不能替代uvrD517am作为抑制子。然而,抑制可能是由于uvrD功能的差异丧失导致的:uvrD517am衍生物中的突变率大幅升高,与uvrD插入突变体中的突变率相当。第二个共抑制突变是编码DNA解旋酶IV的helD基因的一个等位基因,并且可以被helD中的插入突变所替代。第三个共抑制子“srjD”的身份尚不清楚。携带这三个共抑制突变的菌株表现出高重组表型,包括重复序列切除增加。为了解释recJ抑制现象,我们提出抑制子突变导致的抗重组解旋酶活性丧失稳定了在没有recJ的情况下形成的重组中间体。

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