Cloning and Expression Analysis of HAT1 and HDAC1 in the Testes of Mature Yaks and Their Sterile Hybrids

Simple Summary Cattle-yak is the hybrid between male cattle (Bos taurus) and female yak (Bos grunniens). Male cattle-yak can not produce normal sperm. The mechanisms that underlie cattle-yak male sterility have not been elucidated. Histone acetylation is a common regulation mode that plays an important role in the development of gametes. The objective of this study was to explore the molecular mechanism of male sterility in yak hybrids based on histone acetyltransferase 1 (HAT1) and histone deacetylase 1 (HDAC1), two enzymes that regulate histone acetylation. The mRNA and protein expression levels of HAT1 in the testes of adult cattle-yaks were significantly lower than in adult yaks, and the protein expression levels of HDAC1 were significantly higher than in yaks. In addition, H3K9 acetylation levels in cattle-yak testes were significantly lower than in yaks. These results suggest that male sterility in cattle-yaks might be associated with decreased histone acetylation levels in the testes. Abstract The objective of this study was to explore the molecular mechanism of male sterility in yak hybrids based on HAT1 and HDAC1. Total RNA was extracted from the testes of adult yaks (n = 11) and sterile cattle-yaks (n = 11) followed by reverse transcription. The coding sequence (CDS) of yak HAT1 and HDAC1 were obtained by conventional polymerase chain reaction (PCR) and gene cloning. The testicular mRNA and protein levels of HAT1 and HDAC1 in yaks and cattle-yaks were detected by quantitative PCR (qPCR) and Western blotting, respectively, and the histone H3 lysine 9 (H3K9) histone acetylation level in the testes of yaks and cattle-yaks was assayed using enzyme linked immunosorbent assay (ELISA). The results showed that the CDS of HAT1 and HDAC1 were 1242 bp and 1449 bp in length, encoding 413 and 482 amino acids, respectively; yaks had a similar mRNA sequence as cattle in both genes. The testicular mRNA and protein levels of HAT1 of cattle-yaks were significantly lower than those of yaks, and the protein level of HDAC1 was significantly higher than that of yaks. ELISA showed that the acetylation level of testicular H3K9 was significantly lower in yak hybrids than that of yaks. The present results suggest that the decreased level of HAT1 and increased level of HDAC1 may result in the decreased H3K9 acetylation in cattle-yaks and might be associated with their sterility.

[1]  Yucai Zheng,et al.  Cloning and Expression Analysis of Two Kdm Lysine Demethylases in the Testes of Mature Yaks and Their Sterile Hybrids , 2020, Animals : an open access journal from MDPI.

[2]  M. Snyder,et al.  HAT1 Coordinates Histone Production and Acetylation via H4 Promoter Binding. , 2019, Molecular cell.

[3]  Hui Li,et al.  Advances in the relationship between protein acetylation and sperm DNA stability and sperm motility , 2019 .

[4]  Xiangdong Ding,et al.  Species composition and environmental adaptation of indigenous Chinese cattle , 2017, bioRxiv.

[5]  Michael A. Freitas,et al.  Identification of multiple roles for histone acetyltransferase 1 in replication-coupled chromatin assembly , 2017, Nucleic acids research.

[6]  Di Zhang,et al.  Metabolic regulation of gene expression through histone acylations , 2016, Nature Reviews Molecular Cell Biology.

[7]  T. Mipam,et al.  Comparative testis proteome of cattleyak from different developmental stages. , 2017, Animal : an international journal of animal bioscience.

[8]  B. Garcia,et al.  Comprehensive analysis of histone post-translational modifications in mouse and human male germ cells , 2016, Epigenetics & Chromatin.

[9]  Xiaoping Ma,et al.  Comparative iTRAQ proteomics revealed proteins associated with spermatogenic arrest of cattleyak. , 2016, Journal of proteomics.

[10]  Honglin Liu,et al.  Molecular characterization and epigenetic regulation of Mei1 in cattle and cattle-yak. , 2015, Gene.

[11]  Benjamin A Garcia,et al.  Comprehensive Catalog of Currently Documented Histone Modifications. , 2015, Cold Spring Harbor perspectives in biology.

[12]  F. Jin,et al.  The expression of GCN5, HDAC1 and DNMT1 in parthenogenetically activated mouse embryos , 2015, Journal of obstetrics and gynaecology : the journal of the Institute of Obstetrics and Gynaecology.

[13]  D. Endo,et al.  Aberrant levels of histone H3 acetylation induce spermatid anomaly in mouse testis , 2015, Histochemistry and Cell Biology.

[14]  Yaqiu Lin,et al.  Histological evaluation and Prdm9 expression level in the testis of sterile male cattle-yaks , 2014 .

[15]  Honglin Liu,et al.  Differential mRNA expression and promoter methylation status of SYCP3 gene in testes of yaks and cattle-yaks. , 2012, Reproduction in domestic animals = Zuchthygiene.

[16]  M. Bartkuhn,et al.  Presence of histone H3 acetylated at lysine 9 in male germ cells and its distribution pattern in the genome of human spermatozoa. , 2011, Reproduction, fertility, and development.

[17]  Y. Hishikawa,et al.  Immunohistochemical Analysis of Histone H3 Modifications in Germ Cells during Mouse Spermatogenesis , 2011, Acta histochemica et cytochemica.

[18]  D. Cortez,et al.  Analysis of protein dynamics at active, stalled, and collapsed replication forks. , 2011, Genes & development.

[19]  A. Agarwal,et al.  Epigenetics, spermatogenesis and male infertility. , 2011, Mutation research.

[20]  O. Dovey,et al.  Histone deacetylase 1 (HDAC1), but not HDAC2, controls embryonic stem cell differentiation , 2010, Proceedings of the National Academy of Sciences.

[21]  Qifa Li,et al.  [Expression levels of Cdc2 and Cdc25A mRNA in cattle, yak, and cattle-yak testis]. , 2009, Yi chuan = Hereditas.

[22]  C. Boyault,et al.  Regulation of protein turnover by acetyltransferases and deacetylases. , 2008, Biochimie.

[23]  G. Draetta,et al.  Role for Histone Deacetylase 1 in Human Tumor Cell Proliferation , 2007, Molecular and Cellular Biology.

[24]  D. Sterner,et al.  Acetylation of Histones and Transcription-Related Factors , 2000, Microbiology and Molecular Biology Reviews.

[25]  T. Kouzarides.  Histone acetylases and deacetylases in cell proliferation. , 1999, Current opinion in genetics & development.

[26]  Y. Hotta,et al.  Fertility investigations in the F1 hybrid and backcross progeny of cattle (Bos taurus) and yak (B. grunniens) in Mongolia. , 1997, Cytogenetics and cell genetics.