Genetically encoded system to track histone modification in vivo

Post-translational histone modifications play key roles in gene regulation, development and differentiation, but their dynamics in living organisms remain almost completely unknown. To address this problem, we developed a genetically encoded system for tracking histone modifications by generating fl...

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Vydáno v:Scientific reports Ročník 3; číslo 1; s. 2436
Hlavní autoři: Sato, Yuko, Mukai, Masanori, Ueda, Jun, Muraki, Michiko, Stasevich, Timothy J., Horikoshi, Naoki, Kujirai, Tomoya, Kita, Hiroaki, Kimura, Taisuke, Hira, Seiji, Okada, Yasushi, Hayashi-Takanaka, Yoko, Obuse, Chikashi, Kurumizaka, Hitoshi, Kawahara, Atsuo, Yamagata, Kazuo, Nozaki, Naohito, Kimura, Hiroshi
Médium: Journal Article
Jazyk:angličtina
Vydáno: London Nature Publishing Group UK 14.08.2013
Nature Publishing Group
Témata:
631/1647/1888/2249
631/1647/2210/2211
631/1647/245/2186
631/337/100/2285
Acetylation
Amino Acid Sequence
Animals
Antibodies
Cell Line
Chromatin
Drosophila melanogaster - embryology
Drosophila melanogaster - metabolism
Drosophila Proteins - metabolism
Embryogenesis
Embryonic Development
Embryonic growth stage
Euchromatin
Gene regulation
Genetic Techniques
Green Fluorescent Proteins - metabolism
Histone deacetylase
Histones - metabolism
Humanities and Social Sciences
Humans
Intracellular Space - metabolism
Localization
Lysine
Lysine - metabolism
Mice
Molecular Sequence Data
multidisciplinary
Post-translation
Protein Processing, Post-Translational
Science
Single-Chain Antibodies - metabolism
Transcription
Zebrafish
ISSN:2045-2322, 2045-2322
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Popis
Shrnutí:Post-translational histone modifications play key roles in gene regulation, development and differentiation, but their dynamics in living organisms remain almost completely unknown. To address this problem, we developed a genetically encoded system for tracking histone modifications by generating fluorescent m odification-specific int racellular anti bodies (mintbodies) that can be expressed in vivo . To demonstrate, an H3 lysine 9 acetylation specific mintbody (H3K9ac-mintbody) was engineered and stably expressed in human cells. In good agreement with the localization of its target acetylation, H3K9ac-mintbody was enriched in euchromatin and its kinetics measurably changed upon treatment with a histone deacetylase inhibitor. We also generated transgenic fruit fly and zebrafish stably expressing H3K9ac-mintbody for in vivo tracking. Dramatic changes in H3K9ac-mintbody localization during Drosophila embryogenesis could highlight enhanced acetylation at the start of zygotic transcription around mitotic cycle 7. Together, this work demonstrates the broad potential of mintbody and lays the foundation for epigenetic analysis in vivo .
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ISSN:2045-2322
2045-2322
DOI:10.1038/srep02436