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 fluorescent modification-specific intracellular antibodies (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.
1] Graduate School of Frontier Biosciences, Osaka University, Suita. 565-0871, Japan [2] JST, CREST, Suita. 565-0871, Japan.
Yuko
Y
Sato
Masanori
M
Mukai
Jun
J
Ueda
Michiko
M
Muraki
Timothy J
TJ
Stasevich
Naoki
N
Horikoshi
Tomoya
T
Kujirai
Hiroaki
H
Kita
Taisuke
T
Kimura
Seiji
S
Hira
Yasushi
Y
Okada
Yoko
Y
Hayashi-Takanaka
Chikashi
C
Obuse
Hitoshi
H
Kurumizaka
Atsuo
A
Kawahara
Kazuo
K
Yamagata
Naohito
N
Nozaki
Hiroshi
H
Kimura
D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D029721:Drosophila Proteins / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology / Q000378:metabolism; D047108:Embryonic Development; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D057127:Single-Chain Antibodies / Q000378:metabolism; D015027:Zebrafish
D029721:Drosophila Proteins; D006657:Histones; D057127:Single-Chain Antibodies; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008239:Lysine