Linked Life Data

8630261

Source:http://linkedlifedata.com/resource/pubmed/id/8630261

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1996-7-1
pubmed:databankReference
pubmed:abstractText
We have previously described the characterization of genomic clones encoding the entire translated sequence of human pulmonary surfactant protein D (SP-D). We now describe the characterization of a genomic fragment (H5E7) that encodes the entirety of the first translated exon (Exon 2), Intron 1, a short transcribed untranslated sequence (Exon 1; 39 bp), and approximately 4 kb of sequence upstream from the transcription initiation site. The start site was identified by 5'-RACE-PCR cloning and primer extension. A putative TATA box (CATAAATA) was identified approximately 30 bp upstream of the start site. Complete sequencing of a HindIII/SacI fragment (HS-1674) encoding approximately 1.7 kb of sequence 5' to the TATA demonstrated multiple potential cis-regulatory elements including half-site glucocorticoid response elements (GRE), a canonical AP-1 consensus, several AP-1 like sequences, E-box sequences, NF-IL-6 and PEA3 motifs, and putative interferon response elements. H441 lung adenocarcinoma cells, which express low levels of SP-D mRNA, and liver HepG2 cells, were transiently co-transfected with chloramphenicol acetyl transferase (CAT) reporter constructs containing up to 3,000 base pairs of upstream sequence, and with constructs encoding beta-gal. H441 cells transfected with constructs containing at least 161 bp of upstream sequence gave normalized levels of CAT activity greater than or equal to that obtained for parallel positive control transfections using pTK-CAT. Treatment of the cells for 48 h with 50 nM dexamethasone (Dex) gave a 2- to 5-fold increase in CAT activity. Interestingly, a 5'-deletion construct containing 161 bp of upstream sequence (pFS161-CAT) conferred both cell type-restricted and dexamethasone-responsive expression. These studies emphasize the potential complexity of SP-D gene regulation, and further support the hypothesis that the effects of glucocorticoids on SP-D production in vivo are regulated at the level of transcription.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
1044-1549
pubmed:author
pubmed:issnType
Print
pubmed:volume
14
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
121-30
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:8630261-Base Sequence, pubmed-meshheading:8630261-Carrier Proteins, pubmed-meshheading:8630261-Cloning, Molecular, pubmed-meshheading:8630261-Consensus Sequence, pubmed-meshheading:8630261-Gene Deletion, pubmed-meshheading:8630261-Gene Expression Regulation, pubmed-meshheading:8630261-Glucocorticoids, pubmed-meshheading:8630261-Glycoproteins, pubmed-meshheading:8630261-Humans, pubmed-meshheading:8630261-Molecular Sequence Data, pubmed-meshheading:8630261-Mutagenesis, pubmed-meshheading:8630261-Promoter Regions, Genetic, pubmed-meshheading:8630261-Pulmonary Surfactant-Associated Protein D, pubmed-meshheading:8630261-Pulmonary Surfactants, pubmed-meshheading:8630261-Sequence Analysis, DNA, pubmed-meshheading:8630261-TATA Box, pubmed-meshheading:8630261-Transcription, Genetic, pubmed-meshheading:8630261-Transfection
pubmed:year
1996
pubmed:articleTitle
Characterization of the human surfactant protein D promoter: transcriptional regulation of SP-D gene expression by glucocorticoids.
pubmed:affiliation
Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.