A CheZ orthologue in Campylobacter jejuni plays a role in chemotaxis through conserved phosphatase activity-猫眼课题宝

空肠弯曲杆菌中的 CheZ 直系同源物通过保守的磷酸酶活性在趋化性中发挥作用

DOI：

10.1101/2023.01.06.523011

发表时间：

2023

期刊：

影响因子：

通讯作者：

中科院分区：

文献类型：

作者：

Jama A

主要的食源性致病菌Jejunic弯曲杆菌利用趋化运动来定植禽类肠道，也是人类腹泻疾病的致病机制。在Escherichia coliCheY中，CHEZ是一种最初被认为是缺失的磷酸酶。空肠。幽门螺杆菌Hp0170蛋白是CHEZ和ASC的远缘同源蛋白。JejuniCj0700与HP0170同源，Cj0700也可作为弯曲杆菌中的CHEZ同源基因。都是C。JejuniCheV和CheA蛋白还含有一个可能被磷酸化的反应调节(RR)结构域。因此，Cj0700将被预测为去磷酸化C。构建了突变株(Δcj0700)和补体株(Δcj0700，cj0046：：cj0700)。在半固体琼脂上，ΔCj0700突变株较野生型表现出较弱的运动能力，而在互补株中则相反。在下拉试验和细菌双杂交试验中，表达的Cj0700能够与Chey、Chea-RR和CheV相互作用，Cj0700能够去磷酸化Chey和Chea-RR的RR结构域，但效率较低。这些发现证实了Cj0700在Inc.空肠趋血性是通过与Chey相关的磷酸酶活性来实现的，因此很可能是CHEZ的同源基因。Cj0700还部分调节了CheA和Chev上RR结构域的磷酸化水平，尽管这种相互作用的功能后果需要进一步研究。

The major food-borne pathogenCampylobacter jejuniemploys chemotactic motility to colonise the avian gut, and also as a virulence mechanism in human diarrhoeal disease.InEscherichia coliCheY activity is modulated by CheZ, a phosphatase originally thought to be absent inC. jejuni. The Hp0170 protein ofHelicobacter pyloriis a distant homologue of CheZ and, asC. jejuniCj0700 is homologous to HP0170, Cj0700 could also act as a CheZ orthologue inCampylobacter. Both theC. jejuniCheV and CheA proteins also contain a response regulator (RR) domain that may be phosphorylated. Cj0700 would therefore be predicted to dephosphorylateC. jejuniCheY and possibly also the CheV and CheA RR domains.A mutant (Δcj0700)and complement (Δcj0700, cj0046::cj0700) were constructed inC.jejunistrains NCTC11168, NCTC11828 and 81-176. On semisolid agar the Δcj0700mutant strain showed reduced motility relative to wild-type and this phenotype was reversed in the complemented strain. In pull down and bacterial two hybrid assays, expressed Cj0700 was able to interact with CheY, CheA-RR and CheV.Cj0700 is able to dephosphorylate the RR domain of CheY and CheA-RR, but less efficiently, CheV. These findings verify that Cj0700 plays a role inC. jejunichemotaxis through phosphatase activity with respect to CheY, and is hence likely to be a CheZ orthologue. Cj0700 also partially modulates the phosphorylation level of the RR domain on CheA and CheV, although the functional consequences of this interaction require further investigation.