HENMT1 and piRNA Stability Are Required for Adult Male Germ Cell Transposon Repression and to Define the Spermatogenic Program in the Mouse

Shu Ly Lim; Zhi Peng Qu; R Daniel Kortschak; David M Lawrence; Joel Geoghegan; Anna-Lena Hempfling; Martin Bergmann; Christopher C Goodnow; Christopher J Ormandy; Lee Wong; Jeff Mann; Hamish S Scott; Duangporn Jamsai; David L Adelson; Moira K O'Bryan

doi:10.1371/journal.pgen.1005620

HENMT1 and piRNA Stability Are Required for Adult Male Germ Cell Transposon Repression and to Define the Spermatogenic Program in the Mouse

PLoS Genet. 2015 Oct 23;11(10):e1005620. doi: 10.1371/journal.pgen.1005620. eCollection 2015 Oct.

Authors

Affiliations

¹ Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia.
² School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia, Australia.
³ Australian Cancer Research Foundation Cancer Genomics Facility, Centre for Cancer Biology, SA Pathology, Adelaide, South Australia, Australia.
⁴ Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia; Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University Giessen, Giessen, Germany.
⁵ Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University Giessen, Giessen, Germany.
⁶ Australian Phenomics Facility, The Australian National University, Canberra, Australian Capital Territory, Australia.
⁷ The Garvan Institute of Medical Research, Sydney, Darlinghurst, New South Wales, Australia.
⁸ The Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
⁹ Murdoch Childrens Research Institute, The Royal Children's Hospital, Parkville, Victoria, Australia.
¹⁰ School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia, Australia; Australian Cancer Research Foundation Cancer Genomics Facility, Centre for Cancer Biology, SA Pathology, Adelaide, South Australia, Australia; Department of Molecular Pathology, Centre for Cancer Biology, SA Pathology, Adelaide, South Australia, Australia.

Abstract

piRNAs are critical for transposable element (TE) repression and germ cell survival during the early phases of spermatogenesis, however, their role in adult germ cells and the relative importance of piRNA methylation is poorly defined in mammals. Using a mouse model of HEN methyltransferase 1 (HENMT1) loss-of-function, RNA-Seq and a range of RNA assays we show that HENMT1 is required for the 2' O-methylation of mammalian piRNAs. HENMT1 loss leads to piRNA instability, reduced piRNA bulk and length, and ultimately male sterility characterized by a germ cell arrest at the elongating germ cell phase of spermatogenesis. HENMT1 loss-of-function, and the concomitant loss of piRNAs, resulted in TE de-repression in adult meiotic and haploid germ cells, and the precocious, and selective, expression of many haploid-transcripts in meiotic cells. Precocious expression was associated with a more active chromatin state in meiotic cells, elevated levels of DNA damage and a catastrophic deregulation of the haploid germ cell gene expression. Collectively these results define a critical role for HENMT1 and piRNAs in the maintenance of TE repression in adult germ cells and setting the spermatogenic program.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Basic Helix-Loop-Helix Transcription Factors / biosynthesis
Basic Helix-Loop-Helix Transcription Factors / genetics
Chromatin / genetics
DNA Transposable Elements / genetics
Gene Expression Regulation, Developmental
Germ Cells / growth & development
Humans
Infertility, Male / genetics*
Infertility, Male / pathology
Male
Methyltransferases / genetics*
Mice
RNA Stability / genetics*
RNA, Small Interfering / genetics*
Spermatogenesis / genetics*

Substances

Basic Helix-Loop-Helix Transcription Factors
Chromatin
DNA Transposable Elements
Nhlh1 protein, mouse
RNA, Small Interfering
HENMT1 protein, mouse
Methyltransferases

Grants and funding

This work was supported by grants from the Australian Research Council to MKO, HSS, DA and CJO; the New South Wales Cancer Council, Cancer Institute New South Wales, Banque Nationale de Paris-Paribas Australia and New Zealand, RT Hall Trust, and the National Breast Cancer Foundation to CJO, and the Deutsche Forschungsgemeinschaft as part of an International Research Training Group project. MKO, HSS, and CJO are NHMRC Fellows (1058356, 481310, 1023059). CCG is an NHMRC Australia Fellowship. SLL was the recipient of a Lalor Foundation Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.