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Acetylation of Chromatin-Associated Histone H3 Lysine 56 Inhibits the Development of Encysted Artemia Embryos.

Zhou R, Yang F, Chen DF, Sun YX, Yang JS, Yang WJ - PLoS ONE (2013)

Bottom Line: We found that the level of H3K56ac on chromatin increased during diapause formation, and decreased upon diapause termination, remaining basal level throughout subsequent embryonic development.Furthermore, we found that this arrest of the cell cycle and development was induced by H3K56ac and dephosphorylation of the checkpoint protein, retinoblastoma protein.These results have revealed the dynamic change in H3K56ac on chromatin during artemia diapause formation and termination.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Conservation Biology for Endangered Wildlife of the Ministry of Education and College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.

ABSTRACT

Background: As a response to harsh environments, the crustacean artemia produces diapause gastrula embryos (cysts), in which cell division and embryonic development are totally arrested. This dormant state can last for very long periods but be terminated by specific environmental stimuli. Thus, artemia is an ideal model organism in which to study cell cycle arrest and embryonic development.

Principal finding: Our study focuses on the roles of H3K56ac in the arrest of cell cycle and development during artemia diapause formation and termination. We found that the level of H3K56ac on chromatin increased during diapause formation, and decreased upon diapause termination, remaining basal level throughout subsequent embryonic development. In both HeLa cells and artemia, blocking the deacetylation with nicotinamide, a histone deacetylase inhibitor, increased the level of H3K56ac on chromatin and induced an artificial cell cycle arrest. Furthermore, we found that this arrest of the cell cycle and development was induced by H3K56ac and dephosphorylation of the checkpoint protein, retinoblastoma protein.

Conclusions/significance: These results have revealed the dynamic change in H3K56ac on chromatin during artemia diapause formation and termination. Thus, our findings provide insight into the regulation of cell division during arrest of artemia embryonic development and provide further insight into the functions of H3K56ac.

No MeSH data available.


Related in: MedlinePlus

The amount of H3K56ac bound to chromatin decreases during diapause termination and post-diapause development.Diapause embryos, post-diapause embryos and samples during the hatching process were collected as mentioned above. (A) Morphology (upper panel) and BrdU incorporation assay (lower panel) of embryos and nauplius larvae at various developmental stages. The black arrows indicate the representative positive signal. (B) Western blotting analysis. Tubulin was used as a loading control. (C) Western blotting for H3K56ac in total histone extracts. The purity of histone extracts was evaluated by a Coomassie-stained gel. Histone H3 was used as a loading control. (D) Western blotting for H3K56ac in the chromatin and non-chromatin fractions. Coomassie-stained gels showed the protein composition of chromatin and non-chromatin fractions. Tubulin was used as a loading control for non-chromatin fractions. Histone H3 was used as a loading control for chromatin fractions. The intensities of the ECL signals were measured, and the ratio of H3K56ac in the chromatin fraction relative to that in the non-chromatin fraction was calculated (chromatin/non-chromatin) and is shown in the bar graph. The means of three independent biological replicates are shown; error bars represent the S.E.M. (E) Expression levels of Rtt109 and Asf1 by virtual Northern blotting.
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pone-0068374-g003: The amount of H3K56ac bound to chromatin decreases during diapause termination and post-diapause development.Diapause embryos, post-diapause embryos and samples during the hatching process were collected as mentioned above. (A) Morphology (upper panel) and BrdU incorporation assay (lower panel) of embryos and nauplius larvae at various developmental stages. The black arrows indicate the representative positive signal. (B) Western blotting analysis. Tubulin was used as a loading control. (C) Western blotting for H3K56ac in total histone extracts. The purity of histone extracts was evaluated by a Coomassie-stained gel. Histone H3 was used as a loading control. (D) Western blotting for H3K56ac in the chromatin and non-chromatin fractions. Coomassie-stained gels showed the protein composition of chromatin and non-chromatin fractions. Tubulin was used as a loading control for non-chromatin fractions. Histone H3 was used as a loading control for chromatin fractions. The intensities of the ECL signals were measured, and the ratio of H3K56ac in the chromatin fraction relative to that in the non-chromatin fraction was calculated (chromatin/non-chromatin) and is shown in the bar graph. The means of three independent biological replicates are shown; error bars represent the S.E.M. (E) Expression levels of Rtt109 and Asf1 by virtual Northern blotting.

Mentions: Diapause was terminated as described, converting diapause embryos into post-diapause (activated) ones, but no morphological changes were observed during or after this transition. Pre-nauplius emerged from the cyst shell in embryos incubated for 16 to 20 h as described in Materials and Methods. The development of pre-nauplius was completed during the early nauplius stage. The morphological changes at each developmental stage during diapause termination and post-diapause development were observed (Figure 3A).


Acetylation of Chromatin-Associated Histone H3 Lysine 56 Inhibits the Development of Encysted Artemia Embryos.

Zhou R, Yang F, Chen DF, Sun YX, Yang JS, Yang WJ - PLoS ONE (2013)

The amount of H3K56ac bound to chromatin decreases during diapause termination and post-diapause development.Diapause embryos, post-diapause embryos and samples during the hatching process were collected as mentioned above. (A) Morphology (upper panel) and BrdU incorporation assay (lower panel) of embryos and nauplius larvae at various developmental stages. The black arrows indicate the representative positive signal. (B) Western blotting analysis. Tubulin was used as a loading control. (C) Western blotting for H3K56ac in total histone extracts. The purity of histone extracts was evaluated by a Coomassie-stained gel. Histone H3 was used as a loading control. (D) Western blotting for H3K56ac in the chromatin and non-chromatin fractions. Coomassie-stained gels showed the protein composition of chromatin and non-chromatin fractions. Tubulin was used as a loading control for non-chromatin fractions. Histone H3 was used as a loading control for chromatin fractions. The intensities of the ECL signals were measured, and the ratio of H3K56ac in the chromatin fraction relative to that in the non-chromatin fraction was calculated (chromatin/non-chromatin) and is shown in the bar graph. The means of three independent biological replicates are shown; error bars represent the S.E.M. (E) Expression levels of Rtt109 and Asf1 by virtual Northern blotting.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3686719&req=5

pone-0068374-g003: The amount of H3K56ac bound to chromatin decreases during diapause termination and post-diapause development.Diapause embryos, post-diapause embryos and samples during the hatching process were collected as mentioned above. (A) Morphology (upper panel) and BrdU incorporation assay (lower panel) of embryos and nauplius larvae at various developmental stages. The black arrows indicate the representative positive signal. (B) Western blotting analysis. Tubulin was used as a loading control. (C) Western blotting for H3K56ac in total histone extracts. The purity of histone extracts was evaluated by a Coomassie-stained gel. Histone H3 was used as a loading control. (D) Western blotting for H3K56ac in the chromatin and non-chromatin fractions. Coomassie-stained gels showed the protein composition of chromatin and non-chromatin fractions. Tubulin was used as a loading control for non-chromatin fractions. Histone H3 was used as a loading control for chromatin fractions. The intensities of the ECL signals were measured, and the ratio of H3K56ac in the chromatin fraction relative to that in the non-chromatin fraction was calculated (chromatin/non-chromatin) and is shown in the bar graph. The means of three independent biological replicates are shown; error bars represent the S.E.M. (E) Expression levels of Rtt109 and Asf1 by virtual Northern blotting.
Mentions: Diapause was terminated as described, converting diapause embryos into post-diapause (activated) ones, but no morphological changes were observed during or after this transition. Pre-nauplius emerged from the cyst shell in embryos incubated for 16 to 20 h as described in Materials and Methods. The development of pre-nauplius was completed during the early nauplius stage. The morphological changes at each developmental stage during diapause termination and post-diapause development were observed (Figure 3A).

Bottom Line: We found that the level of H3K56ac on chromatin increased during diapause formation, and decreased upon diapause termination, remaining basal level throughout subsequent embryonic development.Furthermore, we found that this arrest of the cell cycle and development was induced by H3K56ac and dephosphorylation of the checkpoint protein, retinoblastoma protein.These results have revealed the dynamic change in H3K56ac on chromatin during artemia diapause formation and termination.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Conservation Biology for Endangered Wildlife of the Ministry of Education and College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.

ABSTRACT

Background: As a response to harsh environments, the crustacean artemia produces diapause gastrula embryos (cysts), in which cell division and embryonic development are totally arrested. This dormant state can last for very long periods but be terminated by specific environmental stimuli. Thus, artemia is an ideal model organism in which to study cell cycle arrest and embryonic development.

Principal finding: Our study focuses on the roles of H3K56ac in the arrest of cell cycle and development during artemia diapause formation and termination. We found that the level of H3K56ac on chromatin increased during diapause formation, and decreased upon diapause termination, remaining basal level throughout subsequent embryonic development. In both HeLa cells and artemia, blocking the deacetylation with nicotinamide, a histone deacetylase inhibitor, increased the level of H3K56ac on chromatin and induced an artificial cell cycle arrest. Furthermore, we found that this arrest of the cell cycle and development was induced by H3K56ac and dephosphorylation of the checkpoint protein, retinoblastoma protein.

Conclusions/significance: These results have revealed the dynamic change in H3K56ac on chromatin during artemia diapause formation and termination. Thus, our findings provide insight into the regulation of cell division during arrest of artemia embryonic development and provide further insight into the functions of H3K56ac.

No MeSH data available.


Related in: MedlinePlus