Specific binding of heterogeneous ribonucleoprotein particle protein K to the human c-myc promoter, in vitro
| Autor: | M, Takimoto, T, Tomonaga, M, Matunis, M, Avigan, H, Krutzsch, G, Dreyfuss, D, Levens |
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| Rok vydání: | 1993 |
| Předmět: |
Cell Nucleus
Base Sequence Heterogeneous Nuclear Ribonucleoprotein A1 Genetic Vectors Immunoblotting Molecular Sequence Data Genes myc RNA-Binding Proteins Oligonucleotides Antisense Transfection Heterogeneous-Nuclear Ribonucleoproteins Peptide Fragments Molecular Weight Oligodeoxyribonucleotides Ribonucleoproteins Heterogeneous-Nuclear Ribonucleoprotein Group A-B Humans Electrophoresis Polyacrylamide Gel Amino Acid Sequence Promoter Regions Genetic HeLa Cells Protein Binding |
| Zdroj: | The Journal of biological chemistry. 268(24) |
| ISSN: | 0021-9258 |
| Popis: | A homopurine/homopyrimidine-like sequence is found 100-150 base pairs upstream of the human c-myc promoter P1. This element, termed the CT-element, has been shown to augment expression from P1, and it serves as a positive transcriptional element when coupled to a heterologous promoter in vivo and in vitro. Synthetic oligonucleotides comprising this element were used to form DNA-protein complexes in electrophoretic mobility shift assays. By using conventional and affinity methods, 61- and 34-kDa proteins were shown to be associated with these complexes. Amino acid sequence analysis and immunological methods have identified these proteins as heterogeneous ribonucleoprotein particle (hnRNP) proteins K and A1. Surprisingly, hnRNP protein K binds to the pyrimidine-rich strand of the CT-element in a sequence-specific manner as well as to the double-stranded molecule. Cotransfection of vectors encoding hnRNP protein K in the sense or anti-sense orientations with reporter plasmids driven by wild-type or mutant CT-elements demonstrates that hnRNP protein K augments gene expression in a cis-element-dependent manner. Taken together, these results suggest that hnRNP protein K may play a role in the transcriptional regulation of the human c-myc gene. |
| Databáze: | OpenAIRE |
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