Inhibitory role of transcription factor COUP-TFII in expression of hTERT in HeLa cells / 中国医学科学杂志(英文版)
Chinese Medical Sciences Journal
;
(4): 157-163, 2004.
Artigo
em Inglês
| WPRIM
| ID: wpr-253999
ABSTRACT
<p><b>OBJECTIVE</b>To clone and identify the proteins involved in regulating the transcription of hTERT and study the role of genes in both hTERT transcription and telomerase activity.</p><p><b>METHODS</b>The full cDNA of COUP-TFII was cloned from HeLa cDNA library by hTERT promoter-based yeast one-hybrid assay and then in-frame inserted into His-tag fusion expression vector pEK318. The His-tag COUP-TFII fusion proteins were purified by Ni-NTA chromatography. The interaction of COUP-TFII with hTERT promoter in vitro was identified by electrophoretic mobility shift assay and Footprint. The role of COUP-TFII in both hTERT transcription and telomerase activity were probed through Luciferase reporter assay, Northern blot, and TRAP-PCR ELISA.</p><p><b>RESULTS</b>COUP-TFII could firmly bind to the downstream E-box and the other two binding sites in hTERT promoter. Luciferase reporter assay indicated COUP-TFII could suppress hTERT promoter activity and stable introduction of COUP-TFII into HeLa cells also decreased both endogenous hTERT transcription and telomerase activity.</p><p><b>CONCLUSION</b>The human COUP-TFII can firmly bind to hTERT promoter, and inhibit telomerase activity through decreasing hTERT transcription. It will greatly facilitate understanding of telomerase regulation in normal and cancer cells.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Farmacologia
/
Fatores de Transcrição
/
Transcrição Gênica
/
Leveduras
/
RNA Mensageiro
/
Células HeLa
/
Receptores de Esteroides
/
Regiões Promotoras Genéticas
/
Clonagem Molecular
/
DNA Complementar
Tipo de estudo:
Estudo prognóstico
Limite:
Humanos
Idioma:
Inglês
Revista:
Chinese Medical Sciences Journal
Ano de publicação:
2004
Tipo de documento:
Artigo
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