HmsP, a putative phosphodiesterase, and HmsT, a putative diguanylate cyclase, control Hms-dependent biofilm formation in Yersinia pestis
HmsP, a putative phosphodiesterase, and HmsT, a putative diguanylate cyclase, control Hms-dependent biofilm formation in Yersinia pestis
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DOI：
10.1111/j.1365-2958.2004.04253.x
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发表时间：
2004-10-01
影响因子：
3.6
通讯作者：
Perry, RD
中科院分区：
文献类型：
--
作者：
Kirillina, O;Fetherston, JD;Perry, RD
鼠疫耶尔森氏菌的Hms（+）表型在低于34 ℃的温度下促进血红素或刚果红（CR）与细胞表面的结合。我们以前证明了Hms（+）表型的温度调节不是在转录水平上控制的。相反，HmsH、HmsR和HmsT在温度从26 ℃变化到37 ℃时降解。我们使用随机转座子诱变，以确定新的基因参与的温度调节表达的HMS表型。其中一个基因，我们指定hmsP，编码一个假定的磷酸二酯酶与保守的EAL基序。在26 ℃和37 ℃下，hmsP的突变引起CR平板上红色菌落的形成。在两种温度下，在质粒上用hmsP（+）补充的菌株形成白色菌落。我们使用结晶紫分析和共聚焦激光扫描显微镜，以证明Hms依赖的生物膜形成的Y。鼠疫细胞Y.鼠疫Hms（+）菌株在26 ℃下生长，但在37 ℃下不生长，在硼硅酸盐玻璃表面上形成生物膜。过表达HmsT（GGDEF结构域蛋白）或在hmsP中具有突变的菌株产生极厚的生物膜。HmsT的每个GGEE残基（氨基酸296-299）以及HmsP的E506和L508残基的丙氨酸取代导致功能丧失。我们认为，HmsT和HmsP共同控制Y中产生的生物膜的量。鼠疫HmsT在37 ℃的降解可能是控制Hms生物膜温度依赖性表达的关键因素。
The Hms(+) phenotype of Yersinia pestis promotes the binding of haemin or Congo red (CR) to the cell surface at temperatures below 34degreesC. We previously demonstrated that temperature regulation of the Hms(+) phenotype is not controlled at the level of transcription. Instead, HmsH, HmsR and HmsT are degraded upon a temperature shift from 26degreesC to 37degreesC. We used random transposon mutagenesis to identify new genes involved in the temperature-regulated expression of the Hms phenotype. One of these genes, which we designated hmsP, encodes a putative phosphodiesterase with a conserved EAL motif. Mutations in hmsP caused formation of red colonies on CR plates at 26degreesC and 37degreesC. Strains complemented with hmsP(+) on a plasmid form white colonies at both temperatures. We used a crystal violet assay and confocal laser scanning microscopy to demonstrate Hms-dependent biofilm formation by Y. pestis cells. Y. pestis Hms(+) strains grown at 26degreesC but not at 37degreesC form a biofilm on borosilicate glass surfaces. Strains that either overexpress HmsT (a GGDEF domain protein) or have a mutation in hmsP produced an extremely thick biofilm. Alanine substitutions for each of the GGEE residues (amino acids 296-299) of HmsT as well as the E506 and L508 residues of HmsP caused a loss of function. We propose that HmsT and HmsP together control the amount of biofilm produced in Y. pestis. Degradation of HmsT at 37degreesC may be a critical factor in controlling the temperature-dependent expression of the Hms biofilm.