Epigenetic Transcriptional Regulation of the Growth Arrest-Specific gene 1 (Gas1) in Hepatic Cell Proliferation at Mononucleosomal Resolution 英文参考文献.docVIP

EpigeneticTranscriptionalRegulationoftheGrowth Arrest-Specificgene1(Gas1)inHepaticCellProliferation atMononucleosomalResolution NataliaSacilotto,AntonioEspert,JosefaCastillo,LuisFranco,GerardoLo′pez-Rodas* ChromatinLaboratory,DepartmentofBiochemistryandMolecularBiology,UniversityofValencia,Burjassot,Valencia,Spain Abstract Background:Gas1(growtharrest-specific1)geneisknowntoinhibitcellproliferationinavarietyofmodels,butitspossible implication in regulating quiescence in adult tissues has not been examined to date. The knowledge of how Gas1 is regulatedinquiescencemaycontributetounderstandthederegulationoccurringinneoplasticdiseases. Methodology/Principal Findings: Gas1 expression has been studied in quiescent murine liver and during the naturally synchronizedcellproliferationafterpartialhepatectomy.Chromatinimmunoprecipitationatnucleosomalresolution(Nuc- ChIP)hasbeenusedtocarryoutthestudypreservingtheinvivoconditions.Transcriptionhasbeenassessedatrealtimeby quantifyingthepresenceofRNApolymeraseIIincodingregions(RNApol-ChIP).IthasbeenfoundthatGas1isexpressed not only in quiescent liver but also at the cell cycle G1/S transition. The latter expression peak had not been previously reported. Two nucleosomes, flanking a nucleosome-free region, are positioned close to the transcription start site. Both nucleosomes slide in going from the active to the inactive state and vice versa. Nuc-ChIP analysis of the acquisition of histone epigenetic marks show distinctive features in both active states: H3K9ac and H3K4me2 are characteristic of transcriptioninG0andH4R3me2inG1/Stransition.Sequential-ChIPanalysisrevealedthatthe‘‘repressing’’markH3K9me2 colocalize with several ‘‘activating’’ marks at nucleosome N-1 when Gas1 is actively transcribed suggesting a greater plasticityofepigeneticmarksthanproposeduntilnow.Therecruitmentofchromatin-remodelingormodifyingcomplexes alsodisplayeddistinctcharacteristicsinquiescenceandtheG1/Stransition. Conclusions/Significance:ThefindingthatGas