Rpb1 Sumoylation in Response to UV Radiation or Transcriptional Impairment in Yeast 英文参考文献.docVIP

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Rpb1 Sumoylation in Response to UV Radiation or Transcriptional Impairment in Yeast 英文参考文献.doc

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Rpb1 Sumoylation in Response to UV Radiation or Transcriptional Impairment in Yeast 英文参考文献

Rpb1SumoylationinResponsetoUVRadiationor TranscriptionalImpairmentinYeast XuefengChen,BaojinDing,DanielleLeJeune,ChristineRuggiero,ShishengLi* DepartmentofComparativeBiomedicalSciences,SchoolofVeterinaryMedicine,LouisianaStateUniversity,BatonRouge,Louisiana,UnitedStatesofAmerica Abstract Covalent modifications of proteins by ubiquitin and the Small Ubiquitin-like MOdifier (SUMO) have been revealed to be involvedinaplethoraofcellularprocesses,includingtranscription,DNArepairandDNAdamageresponses.Ithasbeenwell knownthatinresponsetoDNAdamagethatblockstranscriptionelongation,Rpb1,thelargestsubunitofRNApolymeraseII (PolII),isubiquitylatedandsubsequentlydegradedinmammalianandyeastcells.However,itisstillanenigmaregarding howPolIIrespondstodamagedDNAandconveyssignal(s)forDNAdamage-relatedcellularprocesses.WefoundthatRpb1 is also sumoylated in yeast cells upon UV radiation or impairment of transcription elongation, and this modification is independent of DNA damage checkpoint activation. Ubc9, an E2 SUMO conjugase, and Siz1, an E3 SUMO ligase, play importantrolesinRpb1sumoylation.K1487,whichislocatedintheacidiclinkerregionbetweentheC-terminaldomainand theglobulardomainofRpb1,isthemajorsumoylationsite.Rpb1sumoylationisnotaffectedbyitsubiquitylation,andvice versa, indicating that the two processes do not crosstalk. Abolishment of Rpb1 sumoylation at K1487 does not affect transcription elongation or transcription coupled repair (TCR) of UV-induced DNA damage. However, deficiency in TCR enhancesUV-inducedRpb1sumoylation,presumablyduetothepersistenceoftranscription-blockingDNAlesionsinthe transcribedstrandofagene.Remarkably,abolishmentofRpb1sumoylationatK1487causesenhancedandprolongedUV- induced phosphorylation of Rad53, especially in TCR-deficient cells, suggesting that the sumoylation plays a role in restrainingtheDNAdamagecheckpointresponsecausedbytranscription-blockinglesions.Ourresultsdemonstrateanovel covalent modification of Rpb1 in response to UV induced DNA damage or t