Heavy metal ions in transcription factors from HeLa cells: Sp1, but not octamer transcription factor requires zinc for DNA binding and for activator function
Open Access
- 11 July 1988
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 16 (13), 5771-5781
- https://doi.org/10.1093/nar/16.13.5771
Abstract
Zinc is an important cofactor for many enzymes involved in nucleic acid metabolism such as DNA and RNA polymerases, reverse transcriptase and tRNA synthetases. We have developed an inducible in vitro transcription system using metal-depleted nuclear extracts to reveal the presence and functional relevance of heavy metal ions in transcription factors. Using protein-DNA binding assays (band shift and DNAase I footprint) we show that Sp1, a promoter-specific vertebrate transcription factor that binds to the “GC box” , is reversibly inactivated by metal-depletion. Zinc is required for specific DNA binding in vitro and is also essential for Sp1 factor-directed transcription. In contrast, another factor from HeLa cells, the so-called octamer transcription factor (OTF) that binds to the sequence 5′-ATGCAAATNA, is not affected by metal-depletion and thus seems not to be a zinc metalloprotein.Keywords
This publication has 38 references indexed in Scilit:
- A 100-kD HeLa cell octamer binding protein (OBP100) interacts differently with two separate octamer-related sequences within the SV40 enhancer.Genes & Development, 1987
- A common octamer motif binding protein is involved in the transcription of U6 snRNA by RNA polymerase III and U2 snRNA by RNA polymerase IICell, 1987
- Two zinc fingers of a yeast regulatory protein shown by genetic evidence to be essential for its functionNature, 1987
- Molecular analysis of ouabain-resistant mutants of the mouse lymphoma cell line L5178YMutagenesis, 1987
- A transcription factor which binds to the enhancers of SV40, immunoglobulin heavy chain and U2 snRNA genesNature, 1987
- A lymphoid-specific protein binding to the octamer motif of immunoglobulin genesNature, 1986
- Nuclear factor III, a novel sequence-specific DNA-binding protein from HeLa cells stimulating adenovirus DNA replicationNature, 1986
- An SV40 “enhancer trap” incorporates exogenous enhancers or generates enhancers from its own sequencesCell, 1984
- Tissue Specificity of the Initiation of Immunoglobulin κGene TranscriptionHoppe-Seyler´s Zeitschrift Für Physiologische Chemie, 1984
- Three regions upstream from the cap site are required for efficient and accurate transcription of the rabbit β-globin gene in mouse 3T6 cellsCell, 1983