Characterization of the Promoter Regulatory Region of the Human Pyruvate Dehydrogenase .beta. Gene
| Autor: | Sharon Naik, Mulchand S. Patel, Kunapuli T. Madhusudhan |
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| Rok vydání: | 1995 |
| Předmět: |
Binding Sites
Base Sequence Transcription Genetic Chemistry TATA box Molecular Sequence Data Restriction Mapping Nucleic acid sequence Repressor Pyruvate Dehydrogenase Complex Promoter Pyruvate dehydrogenase phosphatase Biochemistry Molecular biology Gene Expression Regulation Enzymologic DNA-Binding Proteins Chloramphenicol acetyltransferase Consensus Sequence Humans Pyruvate Dehydrogenase (Lipoamide) Promoter Regions Genetic Enhancer Gene Transcription Factors |
| Zdroj: | Biochemistry. 34:1288-1294 |
| ISSN: | 1520-4995 0006-2960 5594-5597 |
| Popis: | A genomic clone (19 kb) harboring the intron-exon sequences and the promoter-regulatory region of the E1 beta gene of human pyruvate dehydrogenase complex was isolated by screening a placental genomic library. The nucleotide sequence of the promoter region (1245 bp) showed 18 differences (including mismatches, insertions, and deletions) as compared to the published sequence [Koike et al. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 5594-5597]. The E1 beta promoter lacked a TATA box homology but contained initiator sequences (two) and Sp1 sites (three) which are frequently found in TATA-less promoters. The DNase I footprinting pattern of the promoter region with crude rat liver nuclear extracts showed at least seven regions of protein binding and nuclease protection (P1-P7). The DNase I protected regions contained consensus nucleotide sequences recognized by GATA-1, Sp1, IgNF-A, Lva, bicoid Q9, NF-kB, HNF-5, H4TF-1, WAP5, and ADH transription factors. Transient expression of chloramphenicol acetyltransferase (CAT) suggested the possible presence of negative elements located within the sequence from -2316 to -930, whereas deletion constructs containing -929 to +32 and -98 to +32 DNA sequences showed approximately 7- and 20-fold increases in CAT activity over the basal CAT activity. Additional studies indicated the presence of an orientation-dependent cis element (or elements) within the region from -282 to -397 that acts as an enhancer or a repressor upon a heterologous thymidine kinase promoter.(ABSTRACT TRUNCATED AT 250 WORDS) |
| Databáze: | OpenAIRE |
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