大肠杆菌全局调控蛋白CsrA中抑制十字花科黑腐病菌胞外蛋白酶活性所必需的氨基酸残基的鉴定  

Identification of the Amino Acid Residue Essential for Inhibition of the Extracellular Protease Activity of Xanthomonas campestris pathovar campestris in the Global Regulator CsrA of Escherichia coli

作　　者：唐东阶[1,2] 钟婉莹[2] 秦春铃[2] 唐纪良[1,2] 晁耐霞[3] 

机构地区：[1]亚热带农业生物资源保护利用国家重点实验室,南宁530005 [2]广西大学生命科学与技术学院,南宁530005 [3]广西医科大学基础医学院,南宁530021

出　　处：《基因组学与应用生物学》2013年第4期440-448,共9页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31071141)资助

摘　　要：CsrA／RsmA蛋白家族是细菌中一类重要的转录后全局调控因子，它们调控细菌的碳代谢、次生代谢、运动、生物被膜形成以及致病等过程。CsrA／RsmA蛋白家族通常由60～72个氨基酸基组成，其中第1至第52位氨基酸高度保守，而第53位以后的氨基酸则高度可变。目前人们认为其氨基酸高度保守性是不同种属细菌CsrA／RsmA功能和作用机理相似的基础，但是其高度可变区与不同种属CsrA／RsmA蛋白功能差异的关系尚不清楚。我们之前的研究显示，大肠杆菌（Escherichiacoli）的CsrA（CsrAE删）蛋白的高度可变区（第53至第61位氨基酸残基）具有强烈的抑制十字花科黑腐病菌（XanthomonztscampeStr／spathovarcampestris，简称‰c）胞外蛋白酶活性的能力。本研究中，我们采用丙氨酸置换方法，确定了CsrA。蒯可变区中抑制xcc胞外蛋白酶活性必需的氨基酸残基。结果显示，CsrAE枷蛋白的第54位赖氨酸（K54）、第60位丝氨酸（S60）和第61位酪氨酸（Y61）置换成丙氨酸后，csrA‰。蛋白丧失了抑制Xcc胞外蛋白酶活性的能力；而其它位点的丙氨酸置换不影响其抑制Xcc胞外蛋白酶活性的能力。这个结果证明K54、S60和Y61是CsrAE.coli抑制Xcc胞外蛋白酶活性所必需的，为揭示大肠杆菌的CsrA。融蛋白抑制Xcc胞外蛋白酶活性的分子机理奠定基础。Members of CsrA/RsmA family of proteins are important global regulators in bacteria, and they control carbon metabolism, secondary metabolism, motility, biofilm formation and pathogenesis at post-transcriptional lev- el. CsrA/RsmA proteins are usually composed of 60-72 amino acids. The amino acids from 1st to 52nd are highly conserved while those after 52＂d are highly variable. It has been known that the highly conservation in amino acid se- quences of CsrAs/RsmAs are the basis for their similarity in biological functions and regulatory mechanisms. How- ever, the function of the variable region remains unclear. In our previous study, we found that the variable region （from 53~ to 61 ~ amino acids） ofEscherichia coli CsrA~ coti has the ability to inhibit the extracellular protease activity ofXanthorrugnas campestris pathovar eampestris （Xcc）. In this study, the amino acid residue essential for inhibition of the extracellular protease activity ofXanthomonas campestris pathovar campestris was determined by using ala- nine substitution analysis. Our results show that K54→A, S60→A and Y61→A substitutions abrogated of the ability ofCsrAE coZi inhibiting the extracellular protease activity of Xcc, indicating that K54, S60 and Y61 are essential for CsrAul to inhibit the extracellular protease activity ofXcc. Our study provide useful information for tmderstanding the molecular mechanism of CsrAE.coti inhibiting the extracellular protease activity ofXcc.

关 键 词：十字花科黒腐病菌 胞外蛋白酶 大肠杆菌 CsrA 丙氨酸置换 

分 类 号：S436.3[农业科学—农业昆虫与害虫防治]