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Initiation of Parental Genome Reprogramming in Fertilized Oocyte by Splicing Kinase SRPK1-Catalyzed Protamine Phosphorylation.

Cell (2020-03-15)
Lan-Tao Gou, Do-Hwan Lim, Wubin Ma, Brandon E Aubol, Yajing Hao, Xin Wang, Jun Zhao, Zhengyu Liang, Changwei Shao, Xuan Zhang, Fan Meng, Hairi Li, Xiaorong Zhang, Ruiming Xu, Dangsheng Li, Michael G Rosenfeld, Pamela L Mellon, Joseph A Adams, Mo-Fang Liu, Xiang-Dong Fu
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The paternal genome undergoes a massive exchange of histone with protamine for compaction into sperm during spermiogenesis. Upon fertilization, this process is potently reversed, which is essential for parental genome reprogramming andย subsequent activation; however, it remains poorly understood how this fundamental process is initiated and regulated. Here, we report that the previously characterized splicing kinase SRPK1 initiates this life-beginning event by catalyzing site-specific phosphorylation of protamine, thereby triggering protamine-to-histone exchange in the fertilized oocyte. Interestingly, protamine undergoes a DNA-dependent phase transition to gel-like condensates and SRPK1-mediated phosphorylation likely helps open up such structures to enhance protamine dismissal by nucleoplasmin (NPM2) and enable the recruitment of HIRA for H3.3 deposition. Remarkably, genome-wide assay for transposase-accessible chromatin sequencing (ATAC-seq) analysis reveals that selective chromatin accessibility in both sperm and MII oocytes is largely erased inย early pronuclei in a protamine phosphorylation-dependent manner, suggesting that SRPK1-catalyzed phosphorylation initiates a highly synchronized reorganization program in both parental genomes.

MATERIALS
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Sigma-Aldrich
Atto 488 Protein Labeling Kit, BioReagent, suitable for fluorescence
Sigma-Aldrich
Aniline Blue solution, 2.5% in 2% acetic acid
Millipore
ANTI-FLAGยฎ antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution