Expression of Ebaf Gene in Rat Oviduct. Evidences of a Regulation of Poly a Site Recognition

MARTIN E. ARGAÑARAZ; PABLO A. VALDECANTOS; CARLOS M. ABATE; DORA C. MICELI

San Carlos de Bariloche, Neuquén

Congreso; XXXVX Reunión Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular - SAIB; 2003

Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular - SAIB

We have previously isolated an oviduct-uterus specific cDNA fragment (Pr14) (Accession No. AF202268) by using RNA arbitrarily primed PCR (RAP-PCR). This EST (Expressed Sequence Tag) corresponds to the 3’ UTR (UnTranslate Region) of the Rattus norvegicus endometrial bleeding-associated factor (ebaf) (Locus Link Accession No. LOC289316) a novel member of the Transforming Growth Factor superfamily. Previous Northern analyses with the Pr14 probe showed two transcripts of about 2.9 kb and 4.1 kb in early pregnant rat oviducts. Analysis of the nucleotide sequence of the 3’ UTR of this gene showed potential poly A signals at 1,028; 1,032; 1,240; 2,957 and 3,344 nucleotides from the end of the CDS (Coding Deducted Sequence) of the terminal exon. Taking into account that the predicted CDS of this gene is 1,101 bp in length, the different theoretical mRNAs could have 2,129; 2,133; 2,341; 4,058 and 4,445 nucleotides respectively. Interestingly, these sizes are in agreement with some of those calculated from the genomic sequence. Our hypothesis is that the 2.9 kb and 4.1 kb transcripts detected in Northern Blots could be originated by an alternative poly A site recognition mechanism. In order to confirm this idea we performed RT-PCR assays using specific primers to amplify the different transcripts. Results obtained are in agreement with our hypothesis.