Linked Life Data

11022037

Source:http://linkedlifedata.com/resource/pubmed/id/11022037

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2001-3-6
pubmed:abstractText
The vasorelaxant and anti-mitogenic activities of the atrial and brain natriuretic peptides depend upon their binding to the type A natriuretic peptide receptor (NPR-A) expressed on the surface of vascular cells. Intervention strategies aimed at controlling NPR-A expression are limited by the paucity of studies in this area. Here we identify a sequence CCAAT between -141 and -137 of the NPR-A promoter that, when mutated, reduces promoter activity by 90% in rat aortic smooth muscle (RASM) cells. Protein/DNA cross-linking and immunoperturbation of electrophoretically shifted complexes formed between RASM nuclear extracts and an oligonucleotide surrounding the CCAAT sequence indicates that the heterotrimeric transcription factor NF-Y binds specifically to the wild-type, but not mutated, CCAAT element. Cotransfection of a dominant negative mutant of the NF-YA subunit results in a concentration-dependent decrease in the activity of the NPR-A promoter in RASM cells confirming that endogenous NF-Y is an activator of the promoter. Mutation of the CCAAT element, in conjunction with mutation of all three Sp1 sites previously shown to be involved in NPR-A promoter regulation, virtually eliminates NPR-A promoter activity in RASM cells. Coexpression of all three NF-Y subunits together with Sp1 in Drosophila cells deficient in these factors indicates that NF-Y and Sp1 act synergistically to reconstitute NPR-A promoter activity. A direct physical association between NF-Y and Sp1 can be demonstrated both in vitro by glutathione S-transferase pull-down assay and in the intact cell by coimmunoprecipitation and functional studies. Together, these studies show that NPR-A promoter activity is dominantly regulated through functional, and possibly physical, interactions of NF-Y and Sp1.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
12
pubmed:volume
276
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1516-22
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:11022037-Animals, pubmed-meshheading:11022037-Aorta, pubmed-meshheading:11022037-Base Sequence, pubmed-meshheading:11022037-Binding Sites, pubmed-meshheading:11022037-CCAAT-Binding Factor, pubmed-meshheading:11022037-Cell Line, pubmed-meshheading:11022037-Cell Nucleus, pubmed-meshheading:11022037-Drosophila, pubmed-meshheading:11022037-Gene Expression Regulation, pubmed-meshheading:11022037-Genes, Reporter, pubmed-meshheading:11022037-Guanylate Cyclase, pubmed-meshheading:11022037-Muscle, Smooth, Vascular, pubmed-meshheading:11022037-Mutagenesis, Site-Directed, pubmed-meshheading:11022037-Promoter Regions, Genetic, pubmed-meshheading:11022037-Protein Subunits, pubmed-meshheading:11022037-Rats, pubmed-meshheading:11022037-Receptors, Atrial Natriuretic Factor, pubmed-meshheading:11022037-Recombinant Fusion Proteins, pubmed-meshheading:11022037-Sp1 Transcription Factor, pubmed-meshheading:11022037-Transcription, Genetic, pubmed-meshheading:11022037-Transfection, pubmed-meshheading:11022037-beta-Galactosidase
pubmed:year
2001
pubmed:articleTitle
Functional interaction of NF-Y and Sp1 is required for type a natriuretic peptide receptor gene transcription.
pubmed:affiliation
Metabolic Research Unit and Department of Medicine, University of California, San Francisco, California 94143-0540, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.