The enzyme telomerase is essential for cell proliferation and tumorigenesis. Telomerase reverse transcriptase (TERT) represents the catalytic subunit of the enzyme. In humans, TERT expression is regulated by several different mechanisms, including alternative splicing. Canis familiaris TERT (dogTERT) has been shown to have a high level of sequence similarity with human TERT, indicating that the dog may represent a suitable animal model for telomerase studies. In the present report we sought to investigate whether dogTERT undergoes alternative splicing. During the analysis of canine mammary tissues (both tumor and paired adjacent to the tumor normal tissues) for dogTERT expression by RT-PCR, we identified eight samples—one tumor and seven adjacent normal—which gave PCR products of unexpected sizes. DNA sequencing revealed two insertions (175 and 28 bp long) and two deletions (17 and 32 bp long), which were encountered in different combinations and gave rise to five different transcripts. The generation of all variants could be explained by the employment of alternative splicing sites within dogTERT genomic sequences. The 175-bp and 28-bp insertions, identified between exons 7 and 8 and between 8 and 9, respectively, constituted unspliced sequences of introns 7 and 8, respectively. Both deletions originated from exon 8 sequence removals due to alternative splicing. All five variants encoded truncated proteins, which lacked essential motifs for reverse transcription and might have thus lost their ability to compose active telomerase enzymes. This is the first identification of alternative splicing events within dogTERT. The results presented here may provide the basis for more thorough studies on the regulation of telomerase activity in canine normal and cancer cells.
Mammalian Genome – Springer Journals
Published: Oct 4, 2008
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