Abstract
The heterogeneous nuclear ribonucleoprotein (hnRNP) K, a component of the hnRNP particles, appears to be involved in several steps of regulation of gene expression. To gain insight into mechanisms of K protein action, we performed two-hybrid screens using full-length hnRNP K as a bait. Several novel protein partners were identified, including Y-box-binding protein (YB-1), splicing factors 9G8 and SRp20, DNA-methyltransferase, hnRNP L, and hnRNP U. In vitro binding studies and co-immunoprecipitation from cellular extracts provided evidence for direct interaction between hnRNP K and YB-1. Two distinct domains in YB-1 were responsible for binding to K protein. Each protein was able to transactivate transcription from a polypyrimidine-rich promoter; however, this effect was reduced when K and YB-1 proteins were coexpressed suggesting a functional interaction between these two proteins.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Binding Sites
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CCAAT-Enhancer-Binding Proteins*
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Cloning, Molecular
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Embryo, Mammalian
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Gene Library
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HeLa Cells
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Heterogeneous-Nuclear Ribonucleoprotein K
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Heterogeneous-Nuclear Ribonucleoprotein L
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Heterogeneous-Nuclear Ribonucleoprotein U
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Heterogeneous-Nuclear Ribonucleoproteins
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Humans
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Jurkat Cells
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Mice
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NFI Transcription Factors
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Nuclear Proteins
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RNA-Binding Proteins / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Ribonucleoproteins / chemistry
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Ribonucleoproteins / genetics
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Ribonucleoproteins / metabolism*
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Transcription Factors / metabolism
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Transfection
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Y-Box-Binding Protein 1
Substances
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CCAAT-Enhancer-Binding Proteins
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DNA-Binding Proteins
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Heterogeneous-Nuclear Ribonucleoprotein K
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Heterogeneous-Nuclear Ribonucleoprotein L
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Heterogeneous-Nuclear Ribonucleoprotein U
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Heterogeneous-Nuclear Ribonucleoproteins
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NFI Transcription Factors
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Nuclear Proteins
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RNA-Binding Proteins
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Recombinant Proteins
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Ribonucleoproteins
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Transcription Factors
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Y-Box-Binding Protein 1
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YBX1 protein, human