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Series GSE119701 Query DataSets for GSE119701
Status Public on Jul 05, 2019
Title PHF7 Links H3K4 Methylation to H2A Ubiquitination During Spermiongenesis [RNA-seq]
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Histone ubiquitination has been suggested to serve as a “tag” for nucleosome removal during histone-to-protamine exchange that is essential for chromatin packaging in round spermatids. Here, we screen for putative E3 ligase and identify that the PHF7, containing both RING finger and PHD domains, is critical for H2A ubiquitination and histone removal. Mechanistically, its PHD domain as a histone code reader can specifically bind H3K4me3/me2 and its RING domain as a histone writer can ubiquinate H2A. PHF7 deficiency results in male infertility in mice by generating Phf7 knockout mice using CRISPR-Cas9 technology. Futhermore, we find impaired histone-to-protamine exchange leads to reduced chromatin compaction in Phf7 knockout sperm. Immunostaining and western blot further confirme abnormal retention of core histones and reduced levels of protamine PRM1 and PRM2 in Phf7 knockout sperm. In order to exclude the possibility that Phf7 regulate the expession of protamine, we sorte the round spermatids from WT and Phf7 knockout females for RNA-seq. The transcriptomes in round spermatids are very similar between WT and Phf7 knockout mice. There are few differentially expressed genes in the two groups and the expression of protamine is also unchanged even if PHF7 deficiency. Therefore we find PHF7 has no effects on gene regulation in histone-to-protamine exchange.
Overall design We isolated round spermatids from testes of WT and Phf7 knockout mice according to reported protocol (Bastos et al., 2005) as different sample for downstream analysis. Round spermatids mRNA profiles of 8-week old WT and Phf7 knockout mice were generated by deep sequencing, in triplicate, using Illumina HiSeq X10.
Contributor(s) Wang X, Li J
Citation(s) 31189663
Submission date Sep 09, 2018
Last update date Oct 05, 2019
Contact name Qing Li
Organization name Chinese Academy of Science
Department institute of biochemistry and cell biology
Lab Jinsong Li's Lab
Street address 320 Yueyang Road
City Shanghai
ZIP/Postal code 200031
Country China
Platforms (1)
GPL21273 HiSeq X Ten (Mus musculus)
Samples (6)
GSM3381036 WT-RS-1
GSM3381037 WT-RS-2
GSM3381038 WT-RS-3
BioProject PRJNA490044
SRA SRP160525

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Supplementary file Size Download File type/resource
GSE119701_RAW.tar 792.0 Mb (http)(custom) TAR (of BW)
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Raw data are available in SRA
Processed data provided as supplementary file

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