Harnessing Natural Diversity to Probe Metabolic Pathways 英文参考文献.docVIP

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Harnessing Natural Diversity to Probe Metabolic Pathways 英文参考文献.doc

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Harnessing Natural Diversity to Probe Metabolic Pathways 英文参考文献

HarnessingNaturalDiversitytoProbe MetabolicPathways Oliver R.Homann1,2,Houjian Cai3,Jeffrey M.Becker3,Susan L.Lindquist2* 1 Committee on Genetics, University of Chicago, Chicago, Illinois, United States of America, 2 Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, UnitedStatesofAmerica,3DepartmentofMicrobiology,UniversityofTennessee,Knoxville,Tennessee,UnitedStatesofAmerica Analyses of cellular processes in the yeast Saccharomyces cerevisiae rely primarily upon a small number of highly domesticated laboratory strains, leaving the extensive natural genetic diversity of the model organism largely unexploredandunexploited.Weaskedifthisdiversitycouldbeusedtoenrichourunderstandingofbasicbiological processes.Asatestcase,weexaminedasimpletrait:theutilizationofdi/tripeptidesasnitrogensources.Thecapacity to import small peptides is likely to be under opposing selective pressures (nutrient utilization versus toxin vulnerability)andmaythereforebesculptedbydiversepathwaysandstrategies.Hitherto,dipeptideutilizationinS. cerevisiae was solely ascribed to the activity of a single protein, the Ptr2p transporter. Using high-throughput phenotyping and several genetically diverse strains, we identified previously unknown cellular activities that contributetothistrait.WefindthattheDal5pallantoate/ureidosuccinatepermeaseisalsocapableoffacilitatingdi/ tripeptidetransport.Moreover,evenintheabsenceofDal5pandPtr2p,anadditionalactivity—almostcertainlythe periplasmicasparaginaseIIAsp3p—facilitatestheutilizationofdipeptideswithC-terminalasparagineresiduesbya differentstrategy.Another,as-yet-unidentifiedactivityenablestheutilizationofdipeptideswithC-terminalarginine residues. The relative contributions of these activities to the utilization of di/tripeptides vary among the strains analyzed, as does the vulnerability of these strains to a toxic dipeptide. Only by sampling the genetic diversity of multiple strains were we able to uncover several previously unrecognized l