title: Structure of the murine macrophage scavenger receptor gene and evaluation of sequences that regulate expression in the macrophage cell line, P388D.
abstract: The structure of the entire murine scavenger receptor gene was determined; it consists of eleven exons spanning more than 60 kilobases. Primer extension showed that transcription initiates at a cluster of sites unassociated with a TATAA element. DNA sequences adjacent to these transcription start sites are highly conserved in murine, human, and bovine genes. When transcriptional activity was tested using a luciferase reporter gene, a promoter fragment (-124 to +20) stimulated luciferase production in P388D1 macrophage-like cells but not in non-macrophage COS-7 or 3T3 cells. A longer promoter fragment (approximately 5 kb) stimulated luciferase activity a further 10-fold in P388D1 cells. However, using a series of fragments from -67 to -1500 bp, a 127 bp fragment (-67 to +50) was as active as a 1500 bp fragment in these assays. Mutation of a putative AP-1 element in the -67 to +50 promoter fragment reduced luciferase activity by 40%; mutation of a putative GATA factor element to TATA increased luciferase activity nearly 2-fold while mutation to AATA had no effect and deletion of the GATA sequence inhibited activity by about 50%. The results suggest that a scavenger receptor promoter fragment can confer cell-specific transcription and that the activity may be mediated in part by factors that recognize the AP-1 and GATA elements.
authors.affiliation: Department of Medicine, Massachusetts General Hospital, Boston 02114, USA.
authors.forename: R P
authors.initials: RP
authors.lastname: Aftring
authors.forename: M W
authors.initials: MW
authors.lastname: Freeman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D003094:Collagen / Q000737:chemistry; D017124:Conserved Sequence; D003593:Cytoplasm / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011971:Receptors, Immunologic / Q000737:chemistry / Q000235:genetics*; D051116:Receptors, Scavenger; D011994:Recombinant Proteins; D012091:Repetitive Sequences, Nucleic Acid; D051117:Scavenger Receptors, Class A
chemical_list: C495140:Msr1 protein, mouse; D011971:Receptors, Immunologic; D051116:Receptors, Scavenger; D011994:Recombinant Proteins; D051117:Scavenger Receptors, Class A; D003094:Collagen; D004247:DNA; D008156:Luciferases