Genome-Scale Analysis of Mycoplasma agalactiae Loci Involved in Interaction with Host Cells 英文参考文献.docVIP

Genome-ScaleAnalysisofMycoplasmaagalactiaeLoci InvolvedinInteractionwithHostCells Agne`sSkapski1,2,Marie-ClaudeHygonenq1,2,EvelineSagne′1,2,Se′bastienGuiral1,2¤,ChristineCitti1,2 , EricBaranowski1,2 * 1INRA,UMR1225,IHAP,Toulouse,France,2Universite′ deToulouse,INP,ENVT,UMR1225,IHAP,Toulouse,France Abstract Mycoplasmaagalactiaeisanimportantpathogenofsmallruminants,inwhichitcausescontagiousagalactia.Itbelongstoa largegroupof‘‘minimalbacteria’’withasmallgenomeandreducedmetaboliccapacitiesthataredependentontheirhost for nutrients. Mycoplasma survival thus relies on intimate contact with host cells, but little is known about the factors involved in these interactions or in the more general infectious process. To address this issue, an assay based on goat epithelialandfibroblasticcellswasusedtoscreenaM.agalactiaeknockoutmutantlibrary.Mutantswithreducedgrowth capacitiesincellculturewereselectedand62genomiclociwereidentifiedascontributingtothisphenotype.Asexpected for minimal bacteria, ‘‘transport and metabolism’’ was the functional category most commonly implicated in this phenotype, but 50% of the selected mutants were disrupted in coding sequences (CDSs) with unknown functions, with surfacelipoproteinsbeingmostcommonlyrepresentedinthiscategory.Sincemycoplasmaslackacellwall,lipoproteinsare likelytobeimportantininteractionswiththehost.Afewintergenicregionswerealsoidentifiedthatmayactasregulatory sequences under co-culture conditions. Interestingly, some mutants mapped to gene clusters that are highly conserved acrossmycoplasmaspeciesbutlocatedindifferentpositions.Oneoftheseclusterswasfoundinatranscriptionallyactive regionoftheM.agalactiaechromosome,downstreamofacrypticpromoter.Apossiblescenariofortheevolutionofthese lociisdiscussed.Finally,severalCDSsidentifiedhereareconservedinotherimportantpathogenicmycoplasmas,andsome were involved in horizontal gene transfer with phylogenetically distant species. These results provide a basis for further decipheringfunctionsmediati