WalK(S221P)突变促进金黄色葡萄球菌荚膜产生的作用研究

郭祖文; 饶一凡; 彭华刚; 饶贤才

doi:10.16016/j.2097-0927.202309117

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WalK(S221P)突变促进金黄色葡萄球菌荚膜产生的作用研究

专题报道：军人团体心理训练研究 | 更新时间：2026-01-09

- WalK(S221P)突变促进金黄色葡萄球菌荚膜产生的作用研究
- WalK(S221P) mutation promotes production of Staphylococcus aureus capsule
- 陆军军医大学学报 2023年45卷第23期页码：2460-2466
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)基础医学院微生物学教研室
  2.400030 重庆，重庆大学生物工程学院
- 作者简介：
  
  [饶贤才，E-mail: raoxiancai@126.com RAO Xiancai, E-mail: raoxiancai@126.com
- 基金信息：
  
  国家自然科学基金面上项目(82272341)
- DOI：10.16016/j.2097-0927.202309117
  中图分类号： R372;R378.11;R394.3
- 收稿：2023-09-20，
  
  修回：2023-10-17，
  
  纸质出版：2023-12-15
- 稿件说明：
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郭祖文, 饶一凡, 彭华刚, 等. WalK(S221P)突变促进金黄色葡萄球菌荚膜产生的作用研究[J]. 陆军军医大学学报, 2023,45(23):2460-2466.

Zuwen GUO, Yifan RAO, Huagang PENG, et al. WalK(S221P) mutation promotes production of Staphylococcus aureus capsule[J]. Journal of Army Medical University, 2023, 45(23): 2460-2466.
郭祖文, 饶一凡, 彭华刚, 等. WalK(S221P)突变促进金黄色葡萄球菌荚膜产生的作用研究[J]. 陆军军医大学学报, 2023,45(23):2460-2466. DOI： 10.16016/j.2097-0927.202309117.

Zuwen GUO, Yifan RAO, Huagang PENG, et al. WalK(S221P) mutation promotes production of Staphylococcus aureus capsule[J]. Journal of Army Medical University, 2023, 45(23): 2460-2466. DOI： 10.16016/j.2097-0927.202309117.

摘要

目的

探究WalK(S221P)突变促进金葡菌荚膜产生的作用及可能机制。

方法

采用转录组测序(RNA-seq)分析WalK(S221P)突变对金葡菌基因表达的影响，采用RT-qPCR检测荚膜合成基因的表达变化。利用荚膜染色和透射电镜观察确认WalK(S221P)突变促进金葡菌荚膜产生的作用，再用RT-qPCR筛选参与介导WalK(S221P)促进荚膜合成的调控分子，通过结合位点分析和凝胶迁移阻滞实验(EMSA)证实WalKR对目标分子的调控作用。

结果

RNA-seq分析显示WalK(S221P)突变对196个金葡菌基因的表达水平影响显著，其中16个荚膜合成操纵子基因在XN108中的表达均显著上调。RT-qPCR检测显示荚膜合成相关

capA

capH

和

capK

在含有WalK(S221P)突变的XN108及K-Newman菌株中分别较回复株XN108-R和野生型Newman菌株显著上调。荚膜染色和透射电镜观察显示XN108的荚膜合成相比于回复株XN108-R明显增多，增厚。RT-qPCR筛选提示WalK(S221P)促进金葡菌荚膜合成可能是通过上调MgrA表达水平实现的，结合位点检索分析及EMSA实验证实WalK激活的WalR具有直接结合

mgrA

基因调控区的功能。

结论

金葡菌WalK(S221P)突变具有促进金葡菌荚膜产生的作用，该作用的发挥可能通过上调MgrA的表达而实现。

Abstract

Objective

To explore the role and possible mechanism of WalK(S221P) mutation in promoting capsule production of

Staphylococcus aureus

(

S. aureus

Methods

RNA sequencing (RNA-seq) was performed to d

etect the effect of WalK(S221P) mutation on gene expression of

S. aureus

strain XN108. RT-qPCR was used to verify the capsule gene variation caused by WalK(S221P) mutation. Capsule staining and transmission electron microscopy (TEM) were employed to determine capsule biosynthesis of

S. aureus

. The possible regulatory genes involved in WalK(S221P) regulation were screened by using RT-qPCR. The binding site of WalKR on the regulatory regions of

mgrA

gene was predicted

and electrophoresis mobility shift assay (EMSA) was conducted to detect the direct binding activity of WalK-activated WalR on the

mgrA

DNA probe.

Results

RNA-seq revealed that the expression of 196 genes altered after WalK(S221P) mutation in

S. aureus

XN108

and among these genes

16 genes involved in capsule biosynthesis were upregulated remarkably. RT-qPCR confirmed the upregulation of

capA

capH

and

capK

in the WalK(S221P)-carried

S. aureus

strains

including XN108 and K-Newman. Capsule staining showed that XN108 had more capsule production than its counterpart XN108-R. TEM observation demonstrated that XN108 had thicker capsule than XN108-R. RT-qPCR indicated an increasing expression of

mgrA

gene in the WalK(S221P)-carried XN108 when compared with that in XN108-R. A putative binding site of WalR on the regulatory regions of

mgrA

gene was predicted

and the direct binding activity of WalK-activated WalR on the

mgrA

regulatory fragment was verified by EMSA.

Conclusion

WalK(S221P) promotes the production of

S. aureus

capsule

and the underlying mechanism might be associated with the upregulation of MgrA

a positive regulator for

S. aureus

capsule biosynthesis.

关键词

Keywords

references

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