Putative cis-Regulatory Elements Associated with Heat Shock Genes Activated During Excystation of Cryptosporidium parvum 英文参考文献.docVIP

Putativecis-RegulatoryElementsAssociatedwithHeat ShockGenesActivatedDuringExcystationof Cryptosporidiumparvum BenjaminCohn1,2.,PatricioManque1,2.¤,AnaM.Lara1,2,MyrnaSerrano1,2,NiharSheth2 ,Gregory Buck1,2 * 1DepartmentofMicrobiologyandImmunology,VirginiaCommonwealthUniversity,Richmond,Virginia,UnitedStatesofAmerica,2CenterfortheStudyofBiological Complexity,VirginiaCommonwealthUniversity,Richmond,Virginia,UnitedStatesofAmerica Abstract Background: Cryptosporidiosis is a ubiquitous infectious disease, caused by the protozoan parasites Cryptosporidium hominis and C. parvum, leading to acute, persistent and chronic diarrhea worldwide. Although the complications of this diseasecanbeserious,evenfatal,inimmunocompromisedpatientsofanyage,theyhavealsobeenfoundtoleadtolong termeffects,includinggrowthinhibitionandimpairedcognitivedevelopment,ininfectedimmunocompetentchildren.The Cryptosporidiumlifecyclealternatesbetweenadormantstage,theoocyst,andahighlyreplicativephasethatincludesboth asexualvegetativestagesaswellassexualstages,implyingfinegeneticregulatorymechanisms.Theparasiteisextremely difficulttostudybecauseitcannotbeculturedinvitroandanimalmodelsareequallychallenging.Therecentpublicationof thegenomesequenceofC.hominisandC.parvumhas,however,significantlyadvancedourunderstandingofthebiology andpathogenesisofthisparasite. Methodology/Principal Findings: Herein, our goal was to identify cis-regulatory elements associated with heat shock responseinCryptosporidiumusingacombinationofinsilicoandrealtimeRT-PCRstrategies.AnalysiswithGibbs-Sampling algorithmsofupstreamnon-translatedregionsoftwelvegenesannotatedasheatshockproteinsintheCryptosporidium genome identified a highly conserved over-represented sequence motif in eleven of them. RT-PCR analyses, described herein and also by others, show that these eleven genes bearing the putative element are induced concurrent with excystationofparasiteoocystsviaheatshock. Conclusions/Significance: Our analyses suggest that occurrences o