Overlapping functions of RBBP4 and RBBP7 in regulating cell proliferation and histone H3.3 deposition during mouse preimplantation development

ABSTRACT

Preimplantation development is critical for reproductive successes in mammals. Thus, it is important to understand how preimplantation embryogenesis is regulated. As a key event of preimplantation development, epigenetic reprogramming has been widely studied, yet how epigenetic complexes regulate preimplantation development remains largely unknown. Retinoblastoma binding protein 4 (RBBP4) and 7 (RBBP7) are integral components of epigenetic complexes including SIN3A, NuRD, and CoREST. Here, we demonstrate that double knockdown of Rbbp4 and 7, but not individually, causes embryonic lethality during the morula-to-blastocyst transition. Mechanistically, depletion of RBBP4 and 7 results in dysregulation of genes related to cell cycle, lineage development, and regulation of transcription, which is accompanied by cell cycle block, disrupted lineage specification and chromatin structure. Interestingly, RBBP4/7 depletion leads to a dramatic increase in H3.3 and H3K27ac abundance during morula-to-blastocyst transition. ChIP-seq analysis in early embryos and embryonic stem cells reveals enrichment of H3.3 at the promoter regions of RBBP4/7 target genes. In summary, our studies demonstrate the compensatory role of RBBP4/7 and reveal its potential mechanisms in preimplantation development.

Summary sentence:

RBBP4 and RBBP7 play a compensatory role in regulating cell proliferation, apoptosis, and histone H3.3 deposition during preimplantation development.

KEYWORDS:

• Embryo
• preimplantation
• epigenetic
• RBBP4
• RBBP7
• HDAC1/2

Acknowledgments

We thank all members of the Zhang laboratory for their helpful discussions.

Supplementary material

Supplemental data for this article can be accessed here

Author contribution statement

K.Z. and L.X. performed study concept and design; X.L. and L.L. performed embryo microinjection and staining, writing, review, and revision of the paper; Y. D. and B.H. performed RNA-seq and ChIP-seq experiments and analysis. S.W. provided technical support and data interpretation. All authors read and approved the final paper.

Ethics statement

All experiments involving lab animals were performed according to the guidelines for the care and use of lab animals and approved by Zhejiang University.

Disclosure statement

We declare that there are no any competing financial interests in relation to the work described.

Data Availability Statement

The datasets generated and/or analysed during the current study are being uploaded to NCBI GEO and will be available soon. https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE188298

Additional information

Funding

The present study was funded by National Natural Science Foundation of China (No. 31872348, No. 31672416, and No. 32072731 to K.Z.; No. 31941007 to L.L. and S.W.), Zhejiang Provincial Natural Science Foundation (Natural Science Foundation of Zhejiang Province LZ21C170001</#AWARD-ID;> to K.Z.) and China Postdoctoral Science Foundation (Postdoctoral Research Foundation of China No. 2020M671742 to L.L.).