Antimicrobial resistance is undoubtedly one of the greatest global health threats.The emergence of multidrug-resistant(MDR)Gram-positive pathogens,such as methicillin-resistant Staphylococcus aureus(MRSA),vancomycin-resistant Enterococcus faecium(VRE),and β-lactamase-resistant Streptococcus pneu-monia,has severely limited our antibiotic arsenal.Numerous ribosome-targeting antibiotics,especially pleuromutilins,oxazolidinones,and streptogramins,are viewed as promising alternatives against aggres-sive MDR pathogens.In this study,we identified a new adenosine triphosphate(ATP)-binding cassete(ABC)-F family determinant,srpA,in Streptococcus suis(S.suis)by means of a comparative analysis of the whole-genome sequences of tiamulin(TIA)-resistant and TIA-sensitive bacteria.Functional cloning confirmed that the deduced gene can mediate cross-resistance to pleuromutilins,lincosamides,and streptogramin A in S.suis and S.aureus.A sequence alignment revealed that SrpA shares the highest amino acid identity with Vga(E)(36％)and shows canonical characteristics of ABC-F family members.In SrpA-ribosome docked compounds,the extended loop region of SrpA approaches the valnemulin-binding pocket in the ribosome peptidyl-transferase center and competes with bound valnemulin.A detailed mutational analysis of the loop residues confirmed that this domain is crucial for SrpA activity,as substitutions or truncations of this region affect the efficiency and specificity of antibiotic resistance.Intracellular antibiotics accumulation indicated that SrpA does not act as an efflux pump,while a ribo-some binding assay supported the protective effects of SrpA on the ribosome by preventing antibiotic binding as well as displacing bound drugs.These findings clarify the mechanisms underlying resistance to ribosomal antibiotics.

Chaoyang Zhang;Lu Liu;Peng Zhang;Jingpo Cui;Xiaoxia Qin;Lichao Ma;Kun Han;Zhanhui Wang;Shaolin Wang;Shuangyang Ding;Zhangqi Shen

College of Veterinary Medicine,China Agricultural University,Beijing 100193,China;Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety and Beijing Laboratory for Food Quality and Safety,China Agricultural University,Beijing 100193,China

工程（英文）

[1]Chaoyang Zhang;Lu Liu;Peng Zhang;Jingpo Cui;Xiaoxia Qin;Lichao Ma;Kun Han;Zhanhui Wang;Shaolin Wang;Shuangyang Ding;Zhangqi Shen-.Characterization of a Novel Gene,srpA,Conferring Resistance to Streptogramin A,Pleuromutilins,and Lincosamides in Streptococcus suis)[J].工程（英文）,2022(02):85-94

srpA,Streptogramin,Pleuromutilins,Lincosamides,pleuromutilins,oxazolidinones,streptogramins,aggres,cassete,lincosamides,streptogramin,SrpA,Vga,valnemulin

Characterization,Novel,Gene,Conferring,Resistance,Streptococcus,suis,Antimicrobial,resistance,undoubtedly,greatest,global,health,threats,emergence,multidrug,resistant,MDR,Gram,positive,pathogens,such,methicillin,Staphylococcus,aureus,MRSA,vancomycin,Enterococcus,faecium,VRE,lactamase,pneu,monia,has,severely,limited,our,arsenal,Numerous,ribosome,targeting,antibiotics,especially,viewed,promising,alternatives,against,sive,this,study,identified,new,adenosine,triphosphate,ATP,binding,ABC,family,determinant,by,means,comparative,analysis,whole,genome,sequences,tiamulin,TIA,sensitive,bacteria,Functional,cloning,confirmed,that,deduced,gene,mediate,cross,alignment,revealed,shares,highest,amino,acid,identity,shows,canonical,characteristics,members,docked,compounds,extended,loop,region,approaches,pocket,peptidyl,transferase,center,competes,bound,detailed,mutational,residues,domain,crucial,activity,substitutions,truncations,affect,efficiency,specificity,Intracellular,accumulation,indicated,does,not,efflux,pump,while,assay,supported,protective,effects,preventing,well,displacing,drugs,These,findings,clarify,mechanisms,underlying,ribosomal

0.549318