Amino Acid Substitutions of MagA in Klebsiella pneumoniae Affect the Biosynthesis of the Capsular Polysaccharide 英文参考文献.docVIP

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Amino Acid Substitutions of MagA in Klebsiella pneumoniae Affect the Biosynthesis of the Capsular Polysaccharide 英文参考文献.doc

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Amino Acid Substitutions of MagA in Klebsiella pneumoniae Affect the Biosynthesis of the Capsular Polysaccharide 英文参考文献

AminoAcidSubstitutionsofMagAinKlebsiella pneumoniaeAffecttheBiosynthesisoftheCapsular Polysaccharide Tzu-LungLin1.,Feng-LingYang2.,An-SueiYang3,Hung-PinPeng3,Tsung-LinLi3,Ming-DawTsai2*, Shih-HsiungWu2*,Jin-TownWang1,4* 1Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan, 2Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, 3GenomicsResearchCenter,AcademiaSinica,Taipei,Taiwan,4DepartmentofInternalMedicine,NationalTaiwanUniversityHospital,Taipei,Taiwan Abstract Mucoviscosity-associated gene A (magA) of Klebsiella pneumoniae contributes to K1 capsular polysaccharide (CPS) biosynthesis.Basedonsequencehomologyandgenealignment,themagAgenehasbeenpredictedtoencodeaWzy-type CPSpolymerase.SequencealignmentwiththeWzy_CandRfaLproteinfamilies(whichcatalyzeCPSorlipopolysaccharide (LPS) biosynthesis) and topological analysis has suggested that eight highly conserved residues, including G308, G310, G334, G337, R290, P305, H323, and N324, were located in a hypothetical loop region. Therefore, we used site-directed mutagenesisto studytherole of these residuesin CPS production,and toobservetheconsequent phenotypes suchas mucoviscosity, serum and phagocytosis resistance, and virulence (as assessed in mice) in pyogenic liver abscess strain NTUH-K2044. Alanine substitutions at R290 or H323 abolished all of these properties. The G308A mutant was severely impairedforthesefunctions.TheG334AmutantremainedmucoidwithdecreasedCPSproduction,butitsvirulencewas significantly reduced in vivo. No phenotypic change was observed for strains harboring magA G310A, G337A, P305A, or N324Amutations.Therefore,R290,G308,H323,andG334arefunctionallyimportantresiduesoftheMagA(Wzy)proteinof K.pneumoniaeNTUH-K2044,capsulartypeK1.TheseaminoacidsarealsolikelytobeimportantforthefunctionofWzyin othercapsulartypesinK.pneumoniaeandotherspeciesbearingWzy_Cfamilyproteins. Citation:LinT-L,YangF-L,YangA-S,PengH-P,LiT-L,etal.(2012)AminoAcidSubstitutionsofMagAinKle