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Titolo:
CHARACTERIZATION OF THE HUMAN SURFACTANT-PROTEIN-D PROMOTER - TRANSCRIPTIONAL REGULATION OF SP-D GENE-EXPRESSION BY GLUCOCORTICOIDS
Autore:
RUST K; BINGLE L; MARIENCHECK W; PERSSON A; CROUCH EC;
Indirizzi:
WASHINGTON UNIV,SCH MED,DEPT PATHOL,216 S KINGSHIGHWAY ST LOUIS MO 63110 WASHINGTON UNIV,SCH MED,DEPT PATHOL ST LOUIS MO 63110 WASHINGTON UNIV,SCH MED,DEPT INTERNAL MED ST LOUIS MO 63110
Titolo Testata:
American journal of respiratory cell and molecular biology
fascicolo: 2, volume: 14, anno: 1996,
pagine: 121 - 130
SICI:
1044-1549(1996)14:2<121:COTHSP>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
INVIVO DEXAMETHASONE TREATMENT; CELL-SPECIFIC EXPRESSION; STEROID-HORMONE ACTION; DNA-BINDING; RAT LUNG; A GENE; CHLORAMPHENICOL ACETYLTRANSFERASE; TRANSGENIC MICE; ELEMENTS; INTERFERENCE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
58
Recensione:
Indirizzi per estratti:
Citazione:
K. Rust et al., "CHARACTERIZATION OF THE HUMAN SURFACTANT-PROTEIN-D PROMOTER - TRANSCRIPTIONAL REGULATION OF SP-D GENE-EXPRESSION BY GLUCOCORTICOIDS", American journal of respiratory cell and molecular biology, 14(2), 1996, pp. 121-130

Abstract

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 similar to 4 kb of sequence upstream from the transcriptioninitiation site. The start site was identified by 5'-RACE-PCR cloningand primer extension. A putative TATA box (CATAAATA) was identified similar to 30 bp upstream of the start site. Complete sequencing of a HindIII/SacI fragment (HS-1674) encoding similar to 1.7 kb of sequence 5' to the TATA demonstrated multiple potential cis-regulatory elementsincluding 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.

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Documento generato il 06/04/20 alle ore 04:17:07