title: Isolation and characterization of a TATA-less promoter for the human beta 3 integrin gene.
abstract: Proper expression of the human platelet fibrinogen receptor is necessary for the maintenance of normal hemostasis. This receptor is formed by the heterodimer alpha IIb beta 3, a prototypic member of the integrin family of adhesive molecules. beta 3 is also expressed in other tissues with alpha v as the vitronectin receptor. It was not possible to study the basis for tissue-specific expression of this gene, because the beta 3 gene promoter had not been isolated previously. We have now isolated a 6.0-kb human genomic DNA fragment containing 2.0 kb of sequence 5' to the beta 3 ATG start codon. This clone also contains sequence encoding the signal peptide of the immature beta 3 protein and 3.0 kb of 3' intronic sequence. Primer extension and RNase protection studies of poly A+ RNA from a human erythroleukemia (HEL) cell line indicated a major transcription start site 30 bp upstream of the ATG start codon. In an orientation-dependent manner, a 584-bp fragment 5' to the start codon promotes expression of the chloramphenicol acetyl transferase (CAT) reporter gene in K562 cells. CAT expression from this beta 3 promoter is fivefold above expression from a "promoter-less" control CAT construct. This beta 3 promoter lacks TATA and CAAT cis-acting elements, but there are two Sp1 sites flanking the transcription start site. Other potential transcription factor binding sites are also identified. Phorbol esters (TPA), which increase beta 3 transcription in K562 cells, stimulated transcription from the 584-bp 5' beta 3 region. The isolation of this beta 3 promoter region should permit a more detailed analysis of its transcriptional regulation.
authors.affiliation: Department of Medicine, Johns Hopkins School of Medicine, Baltimore, MD 21205.
authors.forename: M
authors.initials: M
authors.lastname: Villa-Garcia
authors.forename: L
authors.initials: L
authors.lastname: Li
authors.forename: G
authors.initials: G
authors.lastname: Riely
authors.forename: P F
authors.initials: PF
authors.lastname: Bray
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D006801:Humans; D039661:Integrin beta3; D016023:Integrins / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D016385:TATA Box; D014158:Transcription, Genetic
chemical_list: D039661:Integrin beta3; D016023:Integrins