Cell-specific and hormone-regulated expression of gonadotropin-regulated testicular RNA helicase gene (GRTH/Ddx25) resulting from alternative utilization of translation initiation codons in the rat testis.
Gonadotropin-regulated testicular RNA helicase (GRTH) is a novel DEAD-box protein with ATPase and RNA helicase activities. GRTH gene transcription is stimulated by human chorionic gonadotropin (hCG) via cyclic AMP-induced androgen formation in testicular Leydig cells. In this study, immunocytochemical and Western analyses identified GRTH as a developmentally regulated protein in Leydig cells and in germ cells (pachytene spermatocytes and round spermatids) of the rat testis. Three ATGs with the potential for generation of multiple protein species were identified. Germ cells primarily utilized the 1st ATG codon (+1) and contained major proteins of 61/56 kDa, whereas Leydig cells utilized preferentially the 2nd ATG codon (+ 343) with expression of 48/43 kDa species. A 3rd ATG was weakly utilized and yielded a 33-kDa protein only in germ cells. The increase in GRTH 43-kDa protein in Leydig cells caused by hCG treatment was prevented by the androgen receptor antagonist, flutamide. In round spermatids, hCG caused a significant decrease of 61 kDa species and an induction 48/43 kDa species, whereas no changes were observed in pachytene spermatocytes. Reversal of this hormone-induced switch of expression by flutamide indicated a role of androgen in utilization of the 2nd ATG. These studies have demonstrated a cell-specific and hormone-dependent alternative usage of ATG codons in the testis. They have also revealed that the androgen-dependent transcription of GRTH expression in Leydig cells is accompanied by a marked increase of 43-kDa species. The findings indicate that expression of GRTH proteins is regulated by gonadotropin/androgen at the translational level.
Section on Molecular Endocrinology, Endocrinology and Reproduction Research Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892-4510, USA.
Yi
Y
Sheng
Chon-Hwa
CH
Tsai-Morris
Maria L
ML
Dufau
D000595:Amino Acid Sequence; D000726:Androgen Antagonists / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D003062:Codon; D018387:Codon, Initiator; D053487:DEAD-box RNA Helicases; D018076:DNA, Complementary / Q000378:metabolism; D005485:Flutamide / Q000494:pharmacology; D006062:Gonadotropins / Q000378:metabolism*; D006728:Hormones / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D020365:RNA Helicases / Q000737:chemistry* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013087:Spermatids / Q000378:metabolism; D013090:Spermatocytes / Q000378:metabolism; D013091:Spermatogenesis; D013737:Testis / Q000378:metabolism; D013997:Time Factors; D014018:Tissue Distribution; D014158:Transcription, Genetic; D015854:Up-Regulation
D000726:Androgen Antagonists; D003062:Codon; D018387:Codon, Initiator; D018076:DNA, Complementary; D006062:Gonadotropins; D006728:Hormones; D020033:Protein Isoforms; D012333:RNA, Messenger; D005485:Flutamide; C476584:Ddx25 protein, rat; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases