N6-methyladenosine(m^(6)A)is the most prevalent internal RNA modification,and its regulators include writers,readers and erasers.m^(6)A is under stringent control and takes part in many biological events,but it is not...N6-methyladenosine(m^(6)A)is the most prevalent internal RNA modification,and its regulators include writers,readers and erasers.m^(6)A is under stringent control and takes part in many biological events,but it is not known whether there is an interplay between m^(6)A and glycosylation.Here we investigated an m^(6)A reader,YTHDC1,which has been shown to be recruited to the DNA-RNA hybrid at DNA damage sites and regulate homologous recombination(HR)during DNA damage repair.We found that YTHDC1 is subject to O-linked𝛽-N-acetylglucosamine(O-GlcNAc)modification at Ser396 upon DNA damage,which is pivotal for YTHDC1 chromatin binding and ionization radiation induced focus(IRIF)formation.RNA immunoprecipitation(RIP)and molecular dynamics(MD)simulations indicate that O-GlcNAcylation is vital for YTHDC1 to bind with m^(6)A RNA.Fluorescence recovery after photo bleaching(FRAP)analysis revealed that YTHDC1 O-GlcNAcylation is essential for DNA damage-induced YTHDC1-m^(6)A condensate formation.We further demonstrate that YTHDC1 O-GlcNAcylation promotes HR-mediated DNA damage repair and cell survival,probably through recruitment of Rad51 to the damage sites.We propose that YTHDC1 O-GlcNAcylation is instrumental for HR.展开更多
N^(6)-methyladenosine(m^(6)A)on chromosome-associated regulatory RNAs(carRNAs),including repeat RNAs,plays important roles in tuning the chromatin state and transcription,but the intrinsic mechanism remains unclear.He...N^(6)-methyladenosine(m^(6)A)on chromosome-associated regulatory RNAs(carRNAs),including repeat RNAs,plays important roles in tuning the chromatin state and transcription,but the intrinsic mechanism remains unclear.Here,we report that YTHDC1 plays indispensable roles in the self-renewal and differentiation potency of mouse embryonic stem cells(ESCs),which highly depends on the m^(6)A-binding ability.Ythdcl is required for sufficient rRNA synthesis and repression of the 2-cell(2C)transcriptional program in ESCs,which recapitulates the transcriptome regulation by the LINE1 scaffold.Detailed analyses revealed that YTHDC1 recognizes m^(6)A on LINE1 RNAs in the nucleus and regulates the formation of the LINE1-NCL partnership and the chromatin recruitment of KAP1.Moreover,the establishment of H3K9me3 on 2C-related retrotrans-posons is interrupted in Ythdcl-depleted ESCs and inner cell mass(ICM)cells,which consequently increases the transcriptional activities.Our study reveals a role of m^(6)A in regulating the RNA scaffold,providing a new model for the RNA-chromatin cross-talk.展开更多
基金supported by the National Natural Science Foundation of China(NSFC)(32271285 and 31872720)R&D Program of Beijing Municipal Education Commission(KZ202210028043)+5 种基金supported by NSFC(32071277)Natural Science Foundation of Hebei province(C2021201012)S&T Program of Hebei(216Z2602G)Interdisciplinary Research Program of Natural Science of Hebei University(DXK202006)supported by Beijing National Laboratory for Molecular Sciences(BNLMS202108)the Chinese Academy of Sciences Pioneer Hundred Talents Program.W.Q.is supported by NSFC(32088101).
文摘N6-methyladenosine(m^(6)A)is the most prevalent internal RNA modification,and its regulators include writers,readers and erasers.m^(6)A is under stringent control and takes part in many biological events,but it is not known whether there is an interplay between m^(6)A and glycosylation.Here we investigated an m^(6)A reader,YTHDC1,which has been shown to be recruited to the DNA-RNA hybrid at DNA damage sites and regulate homologous recombination(HR)during DNA damage repair.We found that YTHDC1 is subject to O-linked𝛽-N-acetylglucosamine(O-GlcNAc)modification at Ser396 upon DNA damage,which is pivotal for YTHDC1 chromatin binding and ionization radiation induced focus(IRIF)formation.RNA immunoprecipitation(RIP)and molecular dynamics(MD)simulations indicate that O-GlcNAcylation is vital for YTHDC1 to bind with m^(6)A RNA.Fluorescence recovery after photo bleaching(FRAP)analysis revealed that YTHDC1 O-GlcNAcylation is essential for DNA damage-induced YTHDC1-m^(6)A condensate formation.We further demonstrate that YTHDC1 O-GlcNAcylation promotes HR-mediated DNA damage repair and cell survival,probably through recruitment of Rad51 to the damage sites.We propose that YTHDC1 O-GlcNAcylation is instrumental for HR.
基金This work was supported by the National Key R&D Program of China(2016YFA0100400,2020YFA0113200,2018YFA0108900 and 2016YFC1000600)the National Natural Science Foundation of China(31922022,31771646,82022027,31721003,31970796,31871448 and 31871446)+3 种基金the Shanghai Rising-Star Program(19QA1409600)the Shanghai Municipal Medical and Health Discipline Construction Projects(2017ZZ02015)the Fundamental Research Funds for the Central Universities(1515219049 and 22120200410)the Major Program of the Development Fund for Shanghai Zhangjiang National Innovation Demonstration Zone(ZJ2018-ZD-004).
文摘N^(6)-methyladenosine(m^(6)A)on chromosome-associated regulatory RNAs(carRNAs),including repeat RNAs,plays important roles in tuning the chromatin state and transcription,but the intrinsic mechanism remains unclear.Here,we report that YTHDC1 plays indispensable roles in the self-renewal and differentiation potency of mouse embryonic stem cells(ESCs),which highly depends on the m^(6)A-binding ability.Ythdcl is required for sufficient rRNA synthesis and repression of the 2-cell(2C)transcriptional program in ESCs,which recapitulates the transcriptome regulation by the LINE1 scaffold.Detailed analyses revealed that YTHDC1 recognizes m^(6)A on LINE1 RNAs in the nucleus and regulates the formation of the LINE1-NCL partnership and the chromatin recruitment of KAP1.Moreover,the establishment of H3K9me3 on 2C-related retrotrans-posons is interrupted in Ythdcl-depleted ESCs and inner cell mass(ICM)cells,which consequently increases the transcriptional activities.Our study reveals a role of m^(6)A in regulating the RNA scaffold,providing a new model for the RNA-chromatin cross-talk.
