A genomic region transcribed into a long noncoding RNA interacts with the Prss42/Tessp-2 promoter in spermatocytes during mouse spermatogenesis, and its flanking sequences can function as enhancers.
Spermatogenesis is regulated by many meiotic stage-specific genes, but how they coordinate the many individual processes is not fully understood. The Prss/Tessp gene cluster is located on mouse chromosome 9F2-F3, and the three genes at this site (Prss42/Tessp-2, Prss43/Tessp-3, and Prss44/Tessp-4) are specifically activated during meiosis in pachytene spermatocytes. We searched for DNase I hypersensitive sites (HSs) and long noncoding RNAs (lncRNAs) at the Prss/Tessp locus to elucidate how they are activated. We found eight DNase I HSs, three of which were testis germ cell-specific at or close to the Prss42/Tessp-2 promoter, and a testis-specific lncRNA, lncRNA-HSVIII, that was transcribed from a region adjacent to the Prss42/Tessp-2 gene. lncRNA-HSVIII transcripts localized to nuclei of most pachytene spermatocytes and the cytosol of stage-X pachytene spermatocytes and spermatids. Chromosome conformation capture revealed that the lncRNA-HSVIII locus specifically interacted with the Prss42/Tessp-2 promoter in primary and secondary spermatocytes. A 5.8-kb genome sequence, encompassing the entire lncRNA-HSVIII sequence and its flanking regions, significantly increased Prss42/Tessp-2 promoter activity using a reporter-gene assay, yet this construct did not change lncRNA-HSVIII expression, indicating that the elevated promoter activity was likely through enhancer activity. Indeed, both upstream and downstream regions of the lncRNA-HSVIII sequence significantly increased Prss42/Tessp-2 promoter activity. Our data therefore identified the direct interaction of a genomic region in the lncRNA-HSVIII locus with the Prss42/Tessp-2 promoter in spermatocytes, and suggested that sequences adjacent to the lncRNA function as enhancers for the Prss42/Tessp-2 gene. Mol. Reprod. Dev. 83: 541-557, 2016. © 2016 Wiley Periodicals, Inc.
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
Ryoma
R
Yoneda
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
Yui
Y
Satoh
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.|Faculty of Science, Department of Biological Sciences, Hokkaido University, Sapporo, Japan.
Ikuya
I
Yoshida
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
Shohei
S
Kawamura
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.|Faculty of Science, Department of Biological Sciences, Hokkaido University, Sapporo, Japan.
Tomoya
T
Kotani
Graduate School of Life Science, Hokkaido University, Sapporo, Japan.|Faculty of Science, Department of Biological Sciences, Hokkaido University, Sapporo, Japan.
Atsushi P
AP
Kimura
D000818:Animals; D004742:Enhancer Elements, Genetic; D056426:Genetic Loci; D008297:Male; D051379:Mice; D062085:RNA, Long Noncoding / Q000096:biosynthesis* / Q000235:genetics; D013090:Spermatocytes / Q000166:cytology / Q000378:metabolism*; D013091:Spermatogenesis / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology*; D014357:Trypsin / Q000096:biosynthesis* / Q000235:genetics
D062085:RNA, Long Noncoding; D014357:Trypsin