Pouillot Flavie, Fayolle Corinne, Carniel Elisabeth
Yersinia Research Unit, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris Cedex 15, France.
Microbiology (Reading). 2007 Aug;153(Pt 8):2426-2434. doi: 10.1099/mic.0.2007/005736-0.
Some adenine methyltransferases have been shown not only to protect specific DNA restriction sites from cleavage by a restriction endonuclease, but also to play a role in various bacterial processes and sometimes in bacterial virulence. This study focused on a type I restriction-modification system (designated yrmI) of Y. pseudotuberculosis. This system is composed of three adjacent genes which could potentially encode an N6-adenine DNA methylase (YamA), an enzyme involved in site-specific recognition (YrsA) and a restriction endonuclease (YreA). Screening of 85 isolates of Y. pestis and Y. pseudotuberculosis indicated that the yrmI system has been lost by Y. pestis and that yamA (but not yrsA or yreA) is present in all Y. pseudotuberculosis strains tested, suggesting that it may be important at some stages of the epidemiological cycle of this species. To further investigate the role of yamA in Y. pseudotuberculosis survival, multiplication or virulence, a DeltayamA mutant of Y. pseudotuberculosis IP32953 was constructed by allelic exchange with a kanamycin cassette. The fact that DeltayamA mutants were obtained indicated that this gene is not essential for Y. pseudotuberculosis viability. The IP32953DeltayamA mutant strain grew as well as the wild-type in a rich medium at both 28 degrees C and 37 degrees C. It also grew normally in a chemically defined medium at 28 degrees C, but exhibited a growth defect at 37 degrees C. In contrast to the Dam adenine methyltransferase, a mutation in yamA did not impair the functions of DNA repair or resistance to detergents. However, the DeltayamA mutant exhibited a virulence defect in a mouse model of intragastric infection. The in silico analysis indicated that the chromosomal region carrying the Y. pseudotuberculosis yrmI locus has been replaced in Y. pestis by a horizontally acquired region which potentially encodes another methyltransferase. YamA might thus be dispensable for Y. pestis growth and virulence because this species has acquired another gene fulfilling the same functions.
一些腺嘌呤甲基转移酶不仅能保护特定的DNA限制位点不被限制内切酶切割,还在各种细菌过程中发挥作用,有时还与细菌毒力有关。本研究聚焦于假结核耶尔森菌的一种I型限制修饰系统(命名为yrmI)。该系统由三个相邻基因组成,它们可能分别编码一种N6-腺嘌呤DNA甲基转移酶(YamA)、一种参与位点特异性识别的酶(YrsA)和一种限制内切酶(YreA)。对85株鼠疫耶尔森菌和假结核耶尔森菌的筛选表明,鼠疫耶尔森菌已丢失yrmI系统,而在所检测的所有假结核耶尔森菌菌株中均存在yamA(但yrsA或yreA不存在),这表明它在该物种的流行病学循环的某些阶段可能很重要。为了进一步研究yamA在假结核耶尔森菌存活、增殖或毒力方面的作用,通过与卡那霉素盒进行等位基因交换构建了假结核耶尔森菌IP32953的DeltayamA突变体。获得DeltayamA突变体这一事实表明该基因对假结核耶尔森菌的生存力并非必需。IP32953DeltayamA突变株在28℃和37℃的丰富培养基中生长情况与野生型相同。它在28℃的化学限定培养基中也能正常生长,但在37℃时表现出生长缺陷。与Dam腺嘌呤甲基转移酶不同,yamA突变不会损害DNA修复功能或对去污剂的抗性。然而,DeltayamA突变体在小鼠胃内感染模型中表现出毒力缺陷。计算机分析表明,携带假结核耶尔森菌yrmI基因座的染色体区域在鼠疫耶尔森菌中已被一个水平获得的区域所取代,该区域可能编码另一种甲基转移酶。因此,YamA对于鼠疫耶尔森菌的生长和毒力可能是可有可无的,因为该物种已经获得了另一个具有相同功能的基因。