Genomic organization and transcriptional analysis of gonadotropin-regulated testicular RNA helicase--GRTH/DDX25 gene.
The gonadotropin-regulated testicular RNA helicase (GRTH/DDX25) is a new member of the DEAD-box protein family. Phylogenetic analysis revealed that GRTH is distantly related to other members of the family. GRTH is transcriptionally up-regulated by gonadotropin, displays ATPase and RNA helicase activities, and participates in germ cell development. To understand the regulation of GRTH gene expression, we investigated its structural organization and aspects of basal transcriptional regulation at the promoter domain. The 20-kb mouse GRTH gene contains 12 coding exons and all but one of its conserved helicase motifs are contained within single exons. GRTH is a TATA-less gene with multiple transcriptional start sites (TSS), GC-rich sequences and a promoter located within -205/+63 bp of the gene. Sequences -852/-354 and -501/-354 bp caused 40-60% and >80% inhibition of transcription in expressing and non-expressing cells, respectively. Transcriptional activity was recovered only in expressing cells by the addition of upstream sequences (-1085/-852 bp). Sp1/Sp3 supported basal transcriptional activity in all cell types, while E-box was an activator-binding site only in non-expressing cells. These findings indicate that a differential pattern of transcriptional regulation may be involved in the control of GRTH gene expression in a cell-specific manner.
Tsai-Morris
Chon-Hwa
CH
Section on Molecular Endocrinology, ERRB, National Institute of Child Health and Human Development, National Institutes of Health, Building 49-6A36, 49 Convent Dr., Bethesda, MD 20892-4510, USA. morrisch@mail.nih.gov
Lei
Shi
S
Jiang
Qiong
Q
Sheng
Yi
Y
Dufau
Maria L
ML
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D053487:DEAD-box RNA Helicases; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D024202:Electrophoretic Mobility Shift Assay; D005091:Exons; D005796:Genes / Q000235:genetics*; D007438:Introns; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D010802:Phylogeny; D011401:Promoter Regions, Genetic / Q000235:genetics; D020365:RNA Helicases / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D017422:Sequence Analysis, DNA; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000235:genetics*
D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D004247:DNA; D008156:Luciferases; C487765:Ddx25 protein, mouse; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases