RRC ID 73124
Author Otsuka K, Yang H, Matsubara S, Shiraishi A, Kurihara M, Satake H, Kimura AP.
Title Evidence for a functional role of Start, a long noncoding RNA, in mouse spermatocytes.
Journal PLoS One
Abstract A mouse testis-specific long noncoding RNA (lncRNA), Start, is localized in the cytosol of Leydig cells and in the nucleus of pachytene spermatocytes. We previously showed that Start regulates steroidogenesis through controlling the expression of Star and Hsd3b1 genes in Leydig cells, but its function in germ cells was not known. Here we verified that a spermatocyte-specific protease gene, Prss43/Tessp-3, was downregulated in Start-knockout testes. To investigate the transcriptional regulatory activity of Start in spermatocytes, we first performed a series of reporter gene assays using a thymidine kinase promoter in spermatocyte-derived GC-2spd(ts) cells. A 5.4-kb genome sequence encompassing Start exhibited enhancer activity for this promoter, and the activity was decreased by knockdown of Start. Deletion of the Start promoter and replacement of the Start sequence abolished the enhancer activity and, consistently, the activity was detected in further experiments only when Start was actively transcribed. We then examined whether the Prss43/Tessp-3 gene could be a target of Start. A reporter gene assay demonstrated that the 5.4-kb sequence exhibited enhancer activity for a Prss43/Tessp-3 promoter in GC-2spd(ts) cells and that the activity was significantly decreased by knockdown of Start. These results suggest that Start functions in transcriptional activation of the Prss43/Tessp-3 gene in spermatocytes. Given that Start is presumed to regulate steroidogenic genes at the posttranscriptional level in Leydig cells, the function in spermatocytes is a novel role of Start. These findings provide an insight into multifunctionality of lncRNAs in the testis.
Volume 17(8)
Pages e0273279
Published 2022-1-1
DOI 10.1371/journal.pone.0273279
PII PONE-D-22-11353
PMID 36006924
PMC PMC9409574
MeSH Animals Gene Expression Regulation Male Mice Promoter Regions, Genetic RNA, Long Noncoding* / genetics RNA, Long Noncoding* / metabolism Spermatocytes* / metabolism Testis / metabolism
IF 2.74
DNA material B6N Mouse BAC clone (RDB07573) B6Ng01‐306O15