HENMT1 and piRNA Stability Are Required for Adult Male Germ Cell Transposon Repression and to Define the Spermatogenic Program in the Mouse.
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.
Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia.
Shu Ly
SL
Lim
School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia, Australia.
Zhi Peng
ZP
Qu
School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia, Australia.
R Daniel
RD
Kortschak
Australian Cancer Research Foundation Cancer Genomics Facility, Centre for Cancer Biology, SA Pathology, Adelaide, South Australia, Australia.
David M
DM
Lawrence
Australian Cancer Research Foundation Cancer Genomics Facility, Centre for Cancer Biology, SA Pathology, Adelaide, South Australia, Australia.
Joel
J
Geoghegan
Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia; Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University Giessen, Giessen, Germany.
Anna-Lena
AL
Hempfling
Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University Giessen, Giessen, Germany.
Martin
M
Bergmann
Australian Phenomics Facility, The Australian National University, Canberra, Australian Capital Territory, Australia.
Christopher C
CC
Goodnow
The Garvan Institute of Medical Research, Sydney, Darlinghurst, New South Wales, Australia.
Christopher J
CJ
Ormandy
The Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
Lee
L
Wong
Murdoch Childrens Research Institute, The Royal Children's Hospital, Parkville, Victoria, Australia.
Jeff
J
Mann
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.
Hamish S
HS
Scott
Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia.
Duangporn
D
Jamsai
School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia, Australia.
David L
DL
Adelson
Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia.
Moira K
MK
O'Bryan
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis / Q000235:genetics; D002843:Chromatin / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000254:growth & development; D006801:Humans; D007248:Infertility, Male / Q000235:genetics* / Q000473:pathology; D008297:Male; D008780:Methyltransferases / Q000235:genetics*; D051379:Mice; D020871:RNA Stability / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D013091:Spermatogenesis / Q000235:genetics*
D051792:Basic Helix-Loop-Helix Proteins; D002843:Chromatin; D004251:DNA Transposable Elements; D008780:Methyltransferases; D034741:RNA, Small Interfering; C498272:Nhlh1 protein, mouse; C000607995:HENMT1 protein, mouse