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Titolo:
A role for A/T-rich sequences and Pit-1/GHF-1 in a distal enhancer locatedin the human growth hormone locus control region with preferential pituitary activity in culture and transgenic mice
Autore:
Jin, Y; Surabhi, RM; Fresnoza, A; Lytras, A; Cattini, PA;
Indirizzi:
Univ Manitoba, Dept Physiol, Winnipeg, MB R3E 3J7, Canada Univ Manitoba Winnipeg MB Canada R3E 3J7 ol, Winnipeg, MB R3E 3J7, Canada
Titolo Testata:
MOLECULAR ENDOCRINOLOGY
fascicolo: 8, volume: 13, anno: 1999,
pagine: 1249 - 1266
SICI:
0888-8809(199908)13:8<1249:ARFASA>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
HUMAN CHORIONIC SOMATOMAMMOTROPIN; TISSUE-SPECIFIC EXPRESSION; PROTEIN-BINDING SITES; TRANSCRIPTION FACTOR; PHYSICAL LINKAGE; GENE-EXPRESSION; PROLACTIN GENE; CELL-LINE; MECHANISM; HOMEOBOX;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Cattini, PA Univ Manitoba, Dept Physiol, 730 William Ave, Winnipeg, MB R3E3J7, Canada Univ Manitoba 730 William Ave Winnipeg MB Canada R3E 3J7 anada
Citazione:
Y. Jin et al., "A role for A/T-rich sequences and Pit-1/GHF-1 in a distal enhancer locatedin the human growth hormone locus control region with preferential pituitary activity in culture and transgenic mice", MOL ENDOCR, 13(8), 1999, pp. 1249-1266

Abstract

A region located remotely upstream of the human pituitary GH (GH-N) gene and required for efficient GH-N gene expression in the pituitary of transgenic mice was cloned as a 1.6-kb Bg/II (1.6G) fragment. The 1.6G fragment in the forward or reverse orientation increased -496GH-N promoter activity significantly in pituitary GC and GH3 cells after gene transfer. The 1.6G fragment was also able to stimulate activity from a minimal thymidine kinase (TK) promoter which, unlike -496GH-N, lacked any Pit-1/GHF-1 element. Enhancer activity was localized by deletion analysis to a 203-bp region in the 3'-end of the 1.6G fragment and was characterized by the presence of a diffuse136-bp nuclease-protected site, observed with pituitary (GC) but not nonpituitary (HeLa) cell nuclear protein. A major low-mobility complex was observed by electrophoretic mobility shift assay (EMSA) with GC cell nuclear protein, and the pattern was distinct from that seen with a HeLa cell extract. The nuclease-protected region contains three A/T-rich Pit-1/GHF-1-like elements, and their disruption, in the context of the 203-bp region fused to the TK promoter, reduced enhancer activity significantly in pituitary cells in; culture. A mutation in this region was also shown to decrease enhancer activity in transgenic mice and correlated with a decrease in the 203-bp enhancer region complex observed by EMSA. The participation of Pit-1/GHF-1 inthis complex is indicated by competition studies with Pit-1/GHF-1 elementsand antibodies, and direct binding of Pit-1/GHF-1 to the A/T-rich sequences was shown by EMSA using recombinant protein. These studies link the A/T-rich sequences to the distal enhancer activity associated with the GH locus control region in vitro and in vivo.

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Documento generato il 28/09/20 alle ore 11:10:21