Rat cytochrome P450c17 gene transcription is initiated at different start sites in extraglandular and glandular tissues.
In the male rat, the mRNA of the steroidogenic cytochrome P450c17 is expressed extraglandularly in the stomach, duodenum, kidney and liver, throughout the animal's lifespan, as demonstrated by reverse transcriptase and polymerase chain reaction (RT-PCR) analysis with specific primers. Northern analysis indicated that all tissues, except the kidney, contain high levels of such mRNA, but the relative mobility of liver mRNA is slightly less than that of the testis and other tissues. Thus, we analysed their 5'- and 3'-untranslated terminal regions (UTRs) by means of 5'- and 3'-rapid amplification of cDNA ends (RACE) techniques. All tissues utilised the same polyadenylation site as the testis. In the 5'-UTR of liver mRNA, however, we found a distal transcription start site (TSS) located 252b upstream of that used in testicular P450c17 mRNA, which is placed 41b upstream of the first ATG. The 5'-UTR sequence of liver P450c17 cDNA exactly matched the contiguous upstream untranslated region of the gene, suggesting that alternative splicing was not involved in the synthesis of liver P450c17 transcript. The other tissues used the same TSS present in the testis. Nevertheless, a second TSS located 125b upstream of the first ATG was found in the stomach and duodenum. These results show that the transcriptional regulation of the CYP17 gene in the rat is complex and differs between tissues in the use of TSSs.
Dalla Valle
Luisa
L
Comparative Endocrinology Laboratory, Department of Biology, University of Padova, Via U. Bassi 58/B, Italy. luisadv@mail.bio.unipd.it
Vianello
Silvia
S
Belvedere
Paola
P
Colombo
Lorenzo
L
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D017398:Alternative Splicing; D000818:Animals; D015152:Blotting, Northern; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary; D004386:Duodenum / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic; D007668:Kidney / Q000201:enzymology*; D008099:Liver / Q000201:enzymology*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics* / Q000378:metabolism; D013270:Stomach / Q000201:enzymology*; D013737:Testis / Q000201:enzymology*; D014158:Transcription, Genetic
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D013254:Steroid 17-alpha-Hydroxylase