Genetically encoded system to track histone modification in vivo
- Authors
- Sato, Y., Mukai, M., Ueda, J., Muraki, M., Stasevich, T.J., Horikoshi, N., Kujirai, T., Kita, H., Kimura, T., Hira, S., Okada, Y., Hayashi-Takanaka, Y., Obuse, C., Kurumizaka, H., Kawahara, A., Yamagata, K., Nozaki, N., and Kimura, H.
- ID
- ZDB-PUB-130903-37
- Date
- 2013
- Source
- Scientific Reports 3: 2436 (Journal)
- Registered Authors
- Kawahara, Atsuo
- Keywords
- none
- MeSH Terms
-
- Acetylation
- Amino Acid Sequence
- Animals
- Cell Line
- Drosophila Proteins/metabolism
- Drosophila melanogaster/embryology
- Drosophila melanogaster/metabolism
- Embryonic Development
- Genetic Techniques*
- Green Fluorescent Proteins/metabolism
- Histones/metabolism*
- Humans
- Intracellular Space/metabolism
- Lysine/metabolism
- Mice
- Molecular Sequence Data
- Protein Processing, Post-Translational*
- Single-Chain Antibodies/metabolism
- Zebrafish
- PubMed
- 23942372 Full text @ Sci. Rep.
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.