DOI: 10.5061/dryad.xd2547dv6
title: Palmitoyltransferase ZDHHC19 regulates histone-to-protamine exchange during spermiogenesis in mice

description: Spermiogenesis, the final phase of spermatogenesis, involves replacing
 histones by protamines, a process known as histone-to-protamine exchange,
 which is crucial for chromatin condensation. While this exchange is
 essential, the precise mechanisms remain incompletely understood. In this
 study, we discovered that the palmitoyltransferase ZDHHC19 functions as a
 key regulator of mouse spermiogenesis. Loss of Zdhhc19 leads to male
 infertility and abnormal sperm morphology. Zdhhc19-deficient sperm exhibit
 impaired histone-to-protamine exchange, leading to retention of histones
 and misdistribution of protamines. Similarly, the palmitoyltransferase
 catalytic site mutant Zdhhc19 C142S knock-in mice show reduced fertility,
 sperm abnormalities, and histone retention. Mechanistically, ZDHHC19
 mediates histone H3 palmitoylation at cysteine 110, weakening H3-H4
 interactions and reducing its affinity for DNA. Palmitoylation of H3
 facilitates histone-to-protamine exchange. This study highlights the
 essential role of ZDHHC19's palmitoyl-transferase activity in
 histone-to-protamine exchange during spermiogenesis and underscores the
 broader significance of histone palmitoylation in chromatin remodeling.
description: Haploid spermatids and spermatocytes were sorted by FACS. In
 brief, adult mouse testes were put in PBS at 33°C for collagenase and
 trypsin digestion. After filtering, single germ cells were stained with 5
 μg/ml Hoechst 33342 at 33°C for 40 min. Then, the two populations were
 separated and sorted by FACS (BD, Aria II, Influx) according to signals in
 630/30 and 460/50 excited by 355nm. 630/30<sup>high</sup>
 460/50<sup>high</sup> population represents spermatocytes, and
 630/30<sup>low</sup> 460/50<sup>low</sup>
 population represents haploid spermatids. In haploid spermatids, the
 FSC<sup>high</sup> population represents round spermatids and
 the FSC<sup>low</sup> population represents elongated
 spermatids.
description: # Palmitoyltransferase ZDHHC19 regulates histone-to-protamine exchange
 during spermiogenesis in mice Dataset DOI:
 [10.5061/dryad.xd2547dv6](10.5061/dryad.xd2547dv6) ## Description of the
 data and file structure This dataset contains the ATAC-seq of GC1 cells
 overexpressing Migr1 (vector) or WT-ZDHHC19 protein, and sorted wild-type
 and *Zdhhc19*-/- mouse (10 weeks) haploid spermatids. Raw reads were
 quality-filtered using Trimmomatic with the default parameters. ATAC-seq
 paired-end data were aligned to the reference mouse genome (GRCm38/mm10)
 using the BWA program. MD5 checksums are included for all FASTQ files to
 validate data integrity. MACS2 files are included for Peak Calling
 Results. ### Files and variables #### File: Migr_1_R1.fastq.gz
 **Description:** ATAC-seq of GC1 cells transfected Migr1 (vector) ####
 File: Migr_2_R1.fastq.gz **Description:** ATAC-seq of GC1 cells
 transfected Migr1 (vector) #### File: Migr_1_R2.fastq.gz
 **Description:** ATAC-seq of GC1 cells transfected Migr1 (vector) ####
 File: ZDHC19_1_R1.fastq.gz **Description:** ATAC-seq of GC1 cells
 transfected WT-ZDHHC19 protein #### File: Migr_2_R2.fastq.gz
 **Description:** ATAC-seq of GC1 cells transfected Migr1 (vector) ####
 File: ZDHC19_1_R2.fastq.gz **Description:** ATAC-seq of GC1 cells
 transfected WT-ZDHHC19 protein #### File: ZDHC19_2_R1.fastq.gz
 **Description:** ATAC-seq of GC1 cells transfected WT-ZDHHC19 protein ####
 File: KO_1_R1.fastq.gz **Description:** ATAC-seq of sorted Zdhhc19-/-
 mouse haploid spermatids #### File: Wt_1_R1.fastq.gz
 **Description:** ATAC-seq of sorted wild-type mouse haploid spermatids
 #### File: MD5.txt **Description:** MD5 checksums for validation of all
 FASTQ files #### File: ZDHC19_2_R2.fastq.gz **Description:** ATAC-seq of
 GC1 cells transfected WT-ZDHHC19 protein #### File: KO_1_R2.fastq.gz
 **Description:** ATAC-seq of sorted Zdhhc19-/- mouse haploid spermatids
 #### File: Wt_1_R2.fastq.gz **Description:** ATAC-seq of sorted wild-type
 mouse haploid spermatids #### File: KO_2_R1.fastq.gz
 **Description:** ATAC-seq of sorted Zdhhc19-/- mouse haploid spermatids
 #### File: Wt_2_R1.fastq.gz **Description:** ATAC-seq of sorted wild-type
 mouse haploid spermatids #### File: KO_2_R2.fastq.gz
 **Description:** ATAC-seq of sorted Zdhhc19-/- mouse haploid spermatids
 #### File: Macs2_v1.zip **Description:** MACS2 Peak Calling Results,
 analyzed by MACS2 software. The file named "All_peakNum.xls" is
 the peak count summary for all samples. For each sample, there are two
 file types. 1. .narrowPeak File: This is a peak coordinate file in the
 narrowPeak format. It can be opened and viewed using software like Excel
 or a standard text editor. 2. .bw File: This is a file in the bigWig (bw)
 format. It can be uploaded to visualization tools such as the UCSC Genome
 Browser or IGV (Integrative Genomics Viewer) to visually display the
 distribution of sequencing reads across a reference genome. #### File:
 Wt_2_R2.fastq.gz **Description:** ATAC-seq of sorted wild-type mouse
 haploid spermatids ## Code/software The ATAC-seq data were aligned to the
 reference mouse genome (GRCm38/mm10) using the BWA program. The peak
 calling profiles were analyzed by the MACS2 software, and the heatmaps
 were drawn by the deeptools tool. ## Access information NA