Bacillus cereus Spores Release Alanine that Synergizes with Inosine to Promote Germination 英文参考文献.docVIP

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Bacillus cereus Spores Release Alanine that Synergizes with Inosine to Promote Germination 英文参考文献.doc

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Bacillus cereus Spores Release Alanine that Synergizes with Inosine to Promote Germination 英文参考文献

BacilluscereusSporesReleaseAlaninethatSynergizes withInosinetoPromoteGermination TetyanaDodatko1,MoniqueAkoachere2,StefanM.Muehlbauer3,ForrestHelfrich4,AmberHowerton2, ChristianRoss5,VickiWysocki4,Ju¨rgenBrojatsch3,ErnestoAbel-Santos2* 1Department ofBiochemistry, Albert Einstein College ofMedicine, Bronx, New York, United States ofAmerica, 2DepartmentofChemistry,University ofNevada, Las Vegas,LasVegas,Nevada,UnitedStatesofAmerica,3DepartmentofMicrobiologyandImmunology,AlbertEinsteinCollegeofMedicine,Bronx,NewYork,UnitedStates ofAmerica,4DepartmentofChemistryandBiochemistry,UniversityofArizona,Tucson,Arizona,UnitedStatesofAmerica,5SchoolofLifeScience,UniversityofNevada, LasVegas,LasVegas,Nevada,UnitedStatesofAmerica Abstract Background:Thefirststepofthebacteriallifecycleisthegerminationofbacterialsporesintotheirvegetativeform,which requires the presence of specific nutrients. In contrast to closely related Bacillus anthracis spores, Bacillus cereus spores germinateinthepresenceofasinglegerminant,inosine,yetwithasignificantlagperiod. MethodsandFindings:Wefoundthattheinitiallagperiodofinosine-treatedgerminationofB.cereussporesdisappeared inthepresenceofsupernatantsderivedfromalreadygerminatedspores.Thelagperiodalsodissipatedwheninosinewas supplemented with the co-germinator alanine. In fact, HPLC-based analysis revealed the presence of amino acids in the supernatant of germinated B. cereus spores. The released amino acids included alanine in concentrations sufficient to promoterapidgerminationofinosine-treatedspores.ThealanineracemaseinhibitorD-cycloserineenhancedgermination ofB.cereusspores,presumablybyincreasingtheL-alanineconcentrationinthesupernatant.Moreover,wefoundthatB. cereussporeslackingthegerminationreceptorsgerIandgerQdidnotgerminateandreleaseaminoacidsinthepresenceof inosine. These mutant spores, however, germinated efficiently when inosine was supplemented with alanine. Finally, removalofreleasedaminoacidsinawashoutexperimentabrogatedinosine-mediatedger