Evidence for positive and negative regulation in the promoter of the chicken delta 1-crystallin gene.
We investigated the role of sequences flanking the transcription initiation site of the delta 1-crystallin gene in transient transfection assays of primary embryonic chicken lens epithelial cells or fibroblasts. Varying lengths of the 5' flanking sequence of the delta 1-crystallin gene (containing some untranslated sequence from exon 1) were fused to the bacterial chloramphenicol acetyltransferase (CAT) gene in the pSVOCAT plasmid. A plasmid carrying the bacterial beta-galactosidase gene driven by the Rous sarcoma virus (RSV) promoter was used as an internal control. Standardized results showed that the sequence located between -120 to -43 exhibited strong promoter activity; however, the promoter activity was markedly reduced (20-fold) when the upstream sequence between -603 and -120 was included in the construct. The delta 1-crystallin promoter displayed little lens preference. This upstream sequence did not reduce the activity of the Simian virus 40 (SV40) early promoter (with or without its enhancer) or the Herpes thymidine kinase promoter in transfection tests, indicating some specificity in its effect. Evidence for a delta 1-crystallin negative trans-acting factor was provided by competition experiments. Our data raise the possibility that expression of the delta 1-crystallin gene involves a negative cis-acting transcription element, a speculation which may deserve further attention in view of the gradual decrease in delta-crystallin synthesis in the developing lens.
Borrás
T
T
Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, Maryland 20892.
Peterson
C A
CA
Piatigorsky
J
J
D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D002642:Chick Embryo; D015500:Chloramphenicol O-Acetyltransferase; D003459:Crystallins / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant; D004848:Epithelium / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005784:Gene Amplification; D005786:Gene Expression Regulation; D007908:Lens, Crystalline / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D003459:Crystallins; D004274:DNA, Recombinant; D004247:DNA; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase