Journal
ENDOCRINE JOURNAL
Volume 48, Issue 4, Pages 465-472Publisher
JAPAN ENDOCRINE SOCIETY
DOI: 10.1507/endocrj.48.465
Keywords
estrogen receptor alpha gene; messenger ribonucleic acid; untranslated exon; alternative splicing; rat
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We have recently isolated two untranslated first exons, exon 0N and exon 0S, of rat estrogen receptor alpha (ER alpha) gene from the liver by use of 5 ' -rapid amplification of the cDNA ends (5 ' -RACE) method. In this communication, we further analyzed the 5 ' -untranslated region (UTR) of the ER alpha mRNA in rat anterior hypophysis in order to investigate the existence of the other 5 ' -untranslated exon(s) of rat ER alpha gene. Total RNA from the anterior hypophysis of 8-week-old female Wistar strain male rats was subjected to 5 ' -RACE with antisense primers located in exon 1 of the rat ER alpha gene and one of the positive clones (clone 35) was sequenced. The nucleotide sequence of clone 35 revealed the insertion of a previously unidentified exon (which we termed exon 0T) between exon 0 (the first reported 5 ' -UTR form of rat ER alpha mRNA) and exon 1 of rat ER alpha mRNA. Analysis of rat genomic DNA indicated that exon 0T was located between exon 0 and exon I of rat ER alpha gene. We further investigated the distribution of ER alpha mRNA containing exon 0T in several brain regions and various peripheral tissues of 8-week-old male and female Wistar strain rats by use of reverse transcription-polymerase chain reaction (RT-PCR). The distribution of the ER alpha mRNA (0T-1) was essentially similar to that of ER alpha mRNA in which exon 0 was spliced onto exon 1 reported previously. These results indicate that (1) exon 0T is a novel untranslated exon of rat ERa gene which is located between exon 0 and exon I on rat genomic DNA, (2) exon 0T is inserted between exon 0 and exon 1 of ER alpha mRNA by alternative splicing, and (3) this alternative splicing may occur in tissues where the transcription of ER alpha gene is initiated from exon 0.
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