Serine/threonine protein kinase mediates rifampicin resistance in Brucella melitensis through interacting with ribosomal protein RpsD and affecting antioxidant capacity
Yaqin Yuan,
Wenqing Ning,
Junjie Chen,
Jiquan Li,
Tianqi Xue,
Cuihong An,
Lingling Mao,
Guangzhi Zhang,
Shizhong Zhou,
Jiabo Ding,
Xiaowen Yang,
Jianqiang Ye
Affiliations
Yaqin Yuan
Jiangsu Key Laboratory of Zoonosis, Key Laboratory of Jiangsu Preventive Veterinary Medicine, Key Laboratory for Avian Preventive Medicine of Ministry of Education, Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, China
Wenqing Ning
Key Laboratory of Animal Biosafe Risk Prevention and Control (North), Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China
Junjie Chen
Tongliao Mongolian Medical Hospital (Tongliao Mongolian Medical Research Institute), China Center for Disease Control and Prevention, Institute of Infectious Disease Control and Prevention, Co-construction research base for brucellosis, Tongliao City, China
Jiquan Li
Qinghai Institute for Endemic Disease Prevention and Control, Xining, China
Tianqi Xue
Jiangsu Key Laboratory of Zoonosis, Key Laboratory of Jiangsu Preventive Veterinary Medicine, Key Laboratory for Avian Preventive Medicine of Ministry of Education, Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, China
Cuihong An
Department of Plague and Brucellosis, Shaanxi Center for Disease Control and Prevention, Xi’an, China
Lingling Mao
Liaoning Province Center for Disease Control and Prevention, Shenyang, China
Guangzhi Zhang
Key Laboratory of Animal Biosafe Risk Prevention and Control (North), Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China
Shizhong Zhou
Key Laboratory of Animal Biosafe Risk Prevention and Control (North), Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China
Jiabo Ding
Key Laboratory of Animal Biosafe Risk Prevention and Control (North), Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China
Xiaowen Yang
Key Laboratory of Animal Biosafe Risk Prevention and Control (North), Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China
Jianqiang Ye
Jiangsu Key Laboratory of Zoonosis, Key Laboratory of Jiangsu Preventive Veterinary Medicine, Key Laboratory for Avian Preventive Medicine of Ministry of Education, Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, China
ABSTRACT Brucellosis, a zoonotic disease, has re-emerged in both humans and animals, causing significant economic losses globally. Recently, an increasing number of rifampicin-resistant Brucella strains have been isolated worldwide without detectable mutations in known antibiotic resistance genes. Here, this study identified the deletion of serine/threonine protein kinase (STPK) gene in B. melitensis as an efficient trigger for rifampicin resistance using bioinformatics predictions, a transposon mutant library, and gene mutation strains. Notably, the absence of the STPK could increase the expression of ribosomal proteins and genes involved in sulfur metabolism and reduced glutathione, and decrease NADPH oxidase activity and NADP+/NADPH ratio, which is associated with the antioxidant capacity of B. melitensis. Moreover, co-immunoprecipitation revealed that STPK could efficiently interact with the ribosomal protein RpsD, possibly altering protein translation and riboswitch expression. These findings demonstrate that the STPK gene mediates resistance by regulating sulfur metabolism to counteract the reactive oxygen species induced by rifampicin. Furthermore, the approaches developed in this study provide a platform for screening new resistance genes in Brucella spp., and the identified STPK or its pathway can serve as a potential target for new drug development against rifampicin-resistant Brucella spp.IMPORTANCENew rifampicin resistance gene in Brucella melitensis is identified via bioinformatics predictions and a whole-genome transposon mutant library, new mechanisms of rifampicin resistance in B. melitensis, and new function of serine/threonine protein kinase gene and its interaction proteins.
