Kelleher J E, Raleigh E A
New England Biolabs, Beverly, Massachusetts 01915.
J Bacteriol. 1994 Oct;176(19):5888-96. doi: 10.1128/jb.176.19.5888-5896.1994.
To understand the role of restriction in regulating gene flow in bacterial populations, we would like to understand the regulation of restriction enzyme activity. Several antirestriction (restriction alleviation) systems are known that reduce the activity of type I restriction enzymes like EcoKI in vivo. Most of these do not act on type II or type III enzymes, but little information is available for the unclassified modification-dependent systems, of which there are three in E. coli K-12. Of particular interest are two physiological controls on type I enzymes: EcoKI restriction is reduced 2 to 3 orders of magnitude following DNA damage, and a similar effect is seen constitutively in Dam- cells. We used the behavior of EcoKI as a control for testing the response to UV treatment of the three endogenous modification-dependent restriction systems of K-12, McrA, McrBC, and Mrr. Two of these were also tested for response to Dam status. We find that all four resident restriction systems show reduced activity following UV treatment, but not in a unified fashion; each response was genetically and physiologically distinct. Possible mechanisms are discussed.
为了理解限制作用在调控细菌群体基因流动中的角色,我们需要了解限制酶活性的调控机制。已知有几种抗限制(限制缓解)系统能够在体内降低I型限制酶(如EcoKI)的活性。其中大多数系统对II型或III型酶不起作用,但对于未分类的依赖修饰的系统,相关信息较少,大肠杆菌K-12中存在三种此类系统。特别值得关注的是对I型酶的两种生理调控:DNA损伤后,EcoKI的限制作用降低2至3个数量级,在Dam-细胞中也能持续观察到类似的效应。我们利用EcoKI的行为作为对照,来测试K-12的三种内源性依赖修饰的限制系统(McrA、McrBC和Mrr)对紫外线处理的反应。其中两种系统还测试了对Dam状态的反应。我们发现,所有四种常驻限制系统在紫外线处理后活性均降低,但方式并不统一;每种反应在遗传和生理上都是独特的。文中讨论了可能的机制。