IDENTIFICATION OF A PROMOTER REGION GENERATING SRY CIRCULAR TRANSCRIPTS BOTH IN GERM-CELLS FROM MALE-ADULT MICE AND IN MALE-MOUSE EMBRYONALGONADS

Citation
S. Dolci et al., IDENTIFICATION OF A PROMOTER REGION GENERATING SRY CIRCULAR TRANSCRIPTS BOTH IN GERM-CELLS FROM MALE-ADULT MICE AND IN MALE-MOUSE EMBRYONALGONADS, Biology of reproduction, 57(5), 1997, pp. 1128-1135
Citations number
26
Categorie Soggetti
Reproductive Biology
Journal title
ISSN journal
00063363
Volume
57
Issue
5
Year of publication
1997
Pages
1128 - 1135
Database
ISI
SICI code
0006-3363(1997)57:5<1128:IOAPRG>2.0.ZU;2-8
Abstract
The mouse testis determining gene Sry is expressed in somatic cells of the differentiating male gonad as a linear transcript, encoding a tra nscription factor containing an HMG box. In the adult mouse testis, Sr y expression occurs in meiotic and postmeiotic germ cells. The mouse g enomic Sry locus is characterized by two arms of a large inverted repe at, flanking a unique region that, between an acceptor and a donor spl ice site, contains a single exon encoding the Sry protein. In germ cel ls from the adult mouse testis, Sry RNA is a circular molecule, which is generated by an inverted splicing event that utilizes the above-men tioned splice sites. Thus, a circular exon is spliced out starting fro m a large linear RNA precursor containing both arms of the inverted re peat, which pair and generate a large stem-loop structure. Using rever se transcription-polymerase chain reaction and an RNase protection ass ay, we have now mapped the 5' end of this precursor RNA in the 5' arm of the inverted repeat. Gel mobility shift assay and in vitro transcri ption with nuclear extracts from adult germ cells further confirm that a region immediately 5' upstream of two transcriptional initiation si tes of the precursor RNA contains a promoter sequence in which two con sensus Sry binding sequences are specifically recognized by nuclear fa ctors present in adult germ cells but not in Sertoli cells. We also sh ow that the linear precursor of the Sry circular transcript and its sp licing product are specifically expressed not only in adult germ cells but also in male embryonal gonads between 11.5 and 13.5 days postcoit um, immediately after the expression of the linear transcript starting from the unique region.