Genomic organization and promoter characterization of the gene encoding the human telomerase reverse transcriptase (hTERT).
The enzyme telomerase plays a crucial role in cellular proliferation and tumorigenesis. By adding hexameric repeats to chromosome ends, it prevents telomeric loss and, thus, entry into senescence. Recent data suggest that expression of the human telomerase reverse transcriptase subunit (hTERT) represents the limiting factor for telomerase activity. To gain an insight into the mechanisms regulating hTERT expression, we have determined the complete genomic organization of the hTERT gene and isolated the 5'- and 3'- flanking region. The hTERT gene encompasses more than 37kb and consists of 16 exons. We show that all hTERT insertion and deletion variants described so far most likely result from the usage of alternative splice consensus sequences in intron or exon regions. Furthermore, we identified a new hTERT splice variant. Analysis of the DNA sequence surrounding the putative transcriptional start region revealed a TATA-less promoter located in a CpG island. A promoter fragment spanning the first 1100bp upstream of the initiating ATG start codon exhibited high-level activity in HEK-293 cells. Several consensus binding sites for the transcription factor Sp1 as well as a c-Myc binding site were identified in this promoter region. Altogether, these results provide the basis for more detailed studies on the regulation of telomerase activity in normal and cancer cells, and may lead to the development of new cancer therapies.
Wick
M
M
Bayer AG, Central Research Division, Department of Molecular Biology, D-51368, Leverkusen, Germany. maresa.wick.mw@bayer.ag.de
Zubov
D
D
Hagen
G
G
D017398:Alternative Splicing; D001483:Base Sequence; D002460:Cell Line; D005091:Exons / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D010920:Placenta / Q000201:enzymology; D011401:Promoter Regions, Genetic / Q000235:genetics; D017422:Sequence Analysis, DNA; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
D019098:Telomerase