Identification, molecular cloning, and cellular distribution of the rat homolog of minichromosome maintenance protein 7 (MCM7) in the rat testis.
As part of a program to decipher the rat testicular proteome, we studied spermatogonia and identified numerous proteins including the human homolog of the Minichromosome Maintenance Protein 7 (MCM7). MCM7 has been implicated in DNA replication in various species, but had not been detected in the testis. Here we describe the cellular distribution of MCM7 transcripts and protein, and their testicular ontogenetic expression. The full-length coding region of the rat MCM7 was also characterized. Northern blot analyses showed that MCM7 transcripts are more abundant in the testis than other organs and confirmed the presence of the 2.4 kb MCM7 transcript at all ages studied. Interestingly, two additional transcripts of 3.2 and 1.6 kb were found from 26 days post partum onwards, when spermatocytes and spermatids accumulate within the tubules. This was confirmed in isolated cell types: the three MCM7 transcripts were observed in meiotic and post-meiotic germ cells. The 3.2 kb isoform has an extended 5' untranslated region (UTR) and the 1.6 kb transcript is the result of alternative splicing of five exons. Western blot and immunohistochemistry experiments evidenced abundant MCM7 in proliferating gonocytes and Sertoli cells in the fetal testis. In the adult testis, an intense signal was observed in spermatogonia and primary spermatocytes. We conclude that the Mcm7 is one example of genes that are differently transcribed and translated in somatic and spermatogenetic cells in mammals. Further work is required to determine the roles of MCM7 in spermatogonia and germ lineage.
INSERM, U625, GERHM, Campus de Beaulieu, Rennes F-35042, France.
Emmanuelle
E
Com
Antoine D
AD
Rolland
Myriam
M
Guerrois
Florence
F
Aubry
Bernard
B
Jégou
Virginie
V
Vallet-Erdtmann
Charles
C
Pineau
D000251:Adenosine Triphosphatases / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*; D007150:Immunohistochemistry; D008297:Male; D064168:Minichromosome Maintenance Complex Component 7; D008969:Molecular Sequence Data; D020543:Proteome; D051381:Rats; D017207:Rats, Sprague-Dawley; D017386:Sequence Homology, Amino Acid; D013091:Spermatogenesis; D013093:Spermatogonia / Q000737:chemistry* / Q000166:cytology; D013737:Testis / Q000737:chemistry* / Q000166:cytology
D004268:DNA-Binding Proteins; D020543:Proteome; D000251:Adenosine Triphosphatases; C512202:MCM7 protein, rat; D064168:Minichromosome Maintenance Complex Component 7