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Design of genetically encoded sensors to detect nucleosome ubiquitination in live cells.
Dos Santos Passos, Carolina; Choi, Yun-Seok; Snow, Christopher D; Yao, Tingting; Cohen, Robert E.
Afiliação
  • Dos Santos Passos C; Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO.
  • Choi YS; Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO.
  • Snow CD; Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO.
  • Yao T; Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, CO.
  • Cohen RE; Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO.
J Cell Biol ; 220(4)2021 04 05.
Article em En | MEDLINE | ID: mdl-33570569
Histone posttranslational modifications (PTMs) are dynamic, context-dependent signals that modulate chromatin structure and function. Ubiquitin (Ub) conjugation to different lysines of histones H2A and H2B is used to regulate diverse processes such as gene silencing, transcriptional elongation, and DNA repair. Despite considerable progress made to elucidate the players and mechanisms involved in histone ubiquitination, there remains a lack of tools to monitor these PTMs, especially in live cells. To address this, we combined an avidity-based strategy with in silico approaches to design sensors for specifically ubiquitinated nucleosomes. By linking Ub-binding domains to nucleosome-binding peptides, we engineered proteins that target H2AK13/15Ub and H2BK120Ub with Kd values from 10-8 to 10-6 M; when fused to fluorescent proteins, they work as PTM sensors in cells. The H2AK13/15Ub-specific sensor, employed to monitor signaling from endogenous DNA damage through the cell cycle, identified and differentiated roles for 53BP1 and BARD1 as mediators of this histone PTM.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Histonas / Nucleossomos / Proteínas Supressoras de Tumor / Ubiquitina-Proteína Ligases / Ubiquitinação / Proteína 1 de Ligação à Proteína Supressora de Tumor p53 Limite: Humans Idioma: En Revista: J Cell Biol Ano de publicação: 2021 Tipo de documento: Article País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
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XML
PubMed Links
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Histonas / Nucleossomos / Proteínas Supressoras de Tumor / Ubiquitina-Proteína Ligases / Ubiquitinação / Proteína 1 de Ligação à Proteína Supressora de Tumor p53 Limite: Humans Idioma: En Revista: J Cell Biol Ano de publicação: 2021 Tipo de documento: Article País de publicação: Estados Unidos