Regulation of Clara cell secretory protein gene expression by the CCAAT-binding factor NF-Y.
Analysis of the transcriptional regulation of the Clara cell secretory protein (CCSP) gene has resulted in the characterization of several trans-acting factors that regulate the activity of this gene. However, little is known about negative regulatory elements involved in CCSP gene transcription. Using transient transfections of luciferase reporter constructs driven by various fragments of the Neotomodon CCSP (nCCSP) promoter, we identified an inhibitory region that contains an inverted CCAAT box located -225 to -221 bp upstream of the transcriptional start site. Sequence analysis in a broad region of the nCCSP promoter (-744/+33) identified another potentially important CCAAT motif (-459/-455). Gel shift and supershift assays indicated that the transcription factor NF-Y binds to both CCAAT boxes. Mutation of the CCAAT motif prevented the in vitro binding of NF-Y and led to a significant increase of CCSP promoter activity in both pulmonary (H441) and non-pulmonary (HeLa and MCF-7) cells, suggesting that NF-Y is involved in a negative transcriptional regulation that may potentially contribute to the highly cell-specific expression of the anti-inflammatory CCSP gene.
Acosta
Adriana
A
Research Unit in Reproductive Medicine, Hospital de Ginecobstetricia Luis Castelazo Ayala, Instituto Mexicano del Seguro Social, Mexico, D.F. 01090, Mexico.
Zariñán
Teresa
T
Macías
Héctor
H
Pasapera
Ana María
AM
Pérez-Solis
Marco Allán
MA
Olivares
Aleida
A
Ulloa-Aguirre
Alfredo
A
Gutiérrez-Sagal
Rubén
R
D000818:Animals; D023081:CCAAT-Binding Factor / Q000235:genetics* / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D014598:Uteroglobin / Q000235:genetics / Q000378:metabolism*
D023081:CCAAT-Binding Factor; C556390:Scgb1a1 protein, mouse; D014157:Transcription Factors; C402244:nuclear factor Y; D014598:Uteroglobin