2024-03-29T08:12:14Zhttps://eprints.lib.hokudai.ac.jp/dspace-oai/requestoai:eprints.lib.hokudai.ac.jp:2115/724782023-01-19T03:10:04Zhdl_2115_20058hdl_2115_149Specific binding of heterogeneous ribonucleoprotein particle protein K to the human c-myc promoter, in vitroTakimoto, MasatoTomonaga, TakeshiMatunis, MichaelAvigan, MarkKrutzsch, HenryDreyfuss, GideonLevens, David490Amino Acid SequenceBase SequenceCell Nucleus/metabolismElectrophoresis, Polyacrylamide GelGenes, mycGenetic VectorsHeLa CellsHeterogeneous Nuclear Ribonucleoprotein A1Heterogeneous-Nuclear Ribonucleoprotein Group A-BHeterogeneous-Nuclear RibonucleoproteinsHumansImmunoblottingMolecular Sequence DataMolecular WeightOligodeoxyribonucleotides/metabolismOligonucleotides, Antisense/metabolismPeptide Fragments/isolation & purificationPromoter Regions, GeneticProtein BindingRNA-Binding Proteins/metabolismRibonucleoproteins/isolation & purificationRibonucleoproteins/metabolismTransfectionA 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.American Society for Biochemistry and Molecular BiologyJournal Articleapplication/pdfhttp://hdl.handle.net/2115/72478https://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/72478/1/RNP-K-JBC93.pdf0021-9258The Journal of biological chemistry2682418249182581993-08-25enginfo:pmid/8349701This research was originally published in the Journal of Biological Chemistry. M Takimoto, T Tomonaga, M Matunis, M Avigan, H Krutzsch, G Dreyfuss and D Levens. Specific binding of heterogeneous ribonucleoprotein particle protein K to the human c-myc promoter, in vitro. J. Biol. Chem. 1993; 268:18249-18258. © the American Society for Biochemistry and Molecular Biology.publisher