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Examination of the Deubiquitylation Site Selectivity of USP51 by Using Chemically Synthesized Ubiquitylated Histones.
Ai, Huasong; Guo, Yu; Sun, Demeng; Liu, Sanling; Qi, Yunkun; Guo, Jing; Qu, Qian; Gong, Qingyue; Zhao, Suwen; Li, Jiabin; Liu, Lei.
Afiliación
  • Ai H; Tsinghua-Peking Center for Life Sciences, MOE Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Department of Chemistry, Tsinghua University, Beijing, 100084, P.R. China.
  • Guo Y; iHuman Institute, School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, P.R. China.
  • Sun D; Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, Shanghai, 201210, P.R. China.
  • Liu S; School of Life Sciences, University of Science and Technology of China, Hefei, 230026, P.R. China.
  • Qi Y; School of Life Sciences, University of Science and Technology of China, Hefei, 230026, P.R. China.
  • Guo J; Tsinghua-Peking Center for Life Sciences, MOE Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Department of Chemistry, Tsinghua University, Beijing, 100084, P.R. China.
  • Qu Q; Tsinghua-Peking Center for Life Sciences, MOE Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Department of Chemistry, Tsinghua University, Beijing, 100084, P.R. China.
  • Gong Q; Tsinghua-Peking Center for Life Sciences, MOE Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Department of Chemistry, Tsinghua University, Beijing, 100084, P.R. China.
  • Zhao S; School of Life Sciences, University of Science and Technology of China, Hefei, 230026, P.R. China.
  • Li J; iHuman Institute, School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, P.R. China.
  • Liu L; School of Life Sciences, University of Science and Technology of China, Hefei, 230026, P.R. China.
Chembiochem ; 20(2): 221-229, 2019 01 18.
Article en En | MEDLINE | ID: mdl-30192049
Histone ubiquitylation and deubiquitylation processes and the mechanisms of their regulation are closely relevant to the field of epigenetics. Recently, the deubiquitylating enzyme USP51 was reported to selectively cleave ubiquitylation on histone H2A at K13 or K15 (i.e., H2AK13Ub and H2AK15Ub), but not at K119 (i.e., H2AK119Ub), in nucleosomes in vivo. To elucidate the mechanism for the selectivity of USP51, we constructed structurally well-defined in vitro protein systems with a ubiquitin modification at precise sites. A total chemical protein synthesis procedure was developed, wherein hydrazide-based native chemical ligation was used to efficiently generate five ubiquitylated histones (H2AK13Ub, H2AK15Ub, H2AK119Ub, H2BK34Ub, and H2BK120Ub). These synthetic ubiquitylated histones were assembled into nucleosomes and subjected to in vitro USP51 deubiquitylation assays. Surprisingly, USP51 did not show preference between H2AK13/15Ub and H2AK119Ub, in contrast to previous in vivo observations. Accordingly, an understanding of the selectivity of USP51 may require consideration of other factors, such as alternative pre-existing histone modifications, competitive reader proteins, or different nucleosome quality among the in vivo extraction nucleosome and the in vitro reconstitution one. Further experiments established that USP51 in vitro could deubiquitylate a nucleosome carrying H2BK120Ub, but not H2BK34Ub. Molecular dynamics simulations suggested that USP51-catalyzed hydrolysis of ubiquitylated nucleosomes was affected by steric hindrance of the isopeptide bond.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Histonas / Proteasas Ubiquitina-Específicas Límite: Humans Idioma: En Revista: Chembiochem Asunto de la revista: BIOQUIMICA Año: 2019 Tipo del documento: Article Pais de publicación: Alemania
Texto completo
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Imprimir
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Histonas / Proteasas Ubiquitina-Específicas Límite: Humans Idioma: En Revista: Chembiochem Asunto de la revista: BIOQUIMICA Año: 2019 Tipo del documento: Article Pais de publicación: Alemania