Abstract
Polypyrimidine tract-binding protein (PTB) is involved in pre-mRNA splicing and internal ribosomal entry site (IRES)-dependent translation. In order to identify cellular protein(s) interacting with PTB, we performed a yeast two-hybrid screening. Heterogeneous nuclear ribonucleoprotein L (hnRNP L) was identified as a PTB-binding protein. The interaction between PTB and hnRNP L was confirmed in an in vitro binding assay. Both PTB and hnRNP L were found to localize in the nucleoplasm, excepting the nucleoli, in HeLa cells by the green fluorescent protein (GFP)-fused protein detection method. The N-terminal half of PTB (aa 1-329) and most of hnRNP L (aa 141-558) is required for the interaction between PTB and hnRNP L.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
DNA-Binding Proteins / metabolism*
-
Genes, Reporter / genetics
-
Green Fluorescent Proteins
-
HeLa Cells
-
Heterogeneous-Nuclear Ribonucleoprotein L
-
Heterogeneous-Nuclear Ribonucleoproteins
-
Humans
-
Luminescent Proteins / genetics
-
Microscopy, Fluorescence
-
Nuclear Proteins / metabolism
-
Polypyrimidine Tract-Binding Protein
-
Protein Biosynthesis / genetics
-
RNA Precursors / metabolism
-
RNA Splicing / physiology
-
RNA-Binding Proteins / metabolism*
-
Recombinant Fusion Proteins / metabolism
-
Ribonucleoproteins / metabolism*
-
Sequence Deletion / genetics
-
beta-Galactosidase / genetics
-
beta-Galactosidase / metabolism
Substances
-
DNA-Binding Proteins
-
Heterogeneous-Nuclear Ribonucleoprotein L
-
Heterogeneous-Nuclear Ribonucleoproteins
-
Luminescent Proteins
-
Nuclear Proteins
-
RNA Precursors
-
RNA-Binding Proteins
-
Recombinant Fusion Proteins
-
Ribonucleoproteins
-
Polypyrimidine Tract-Binding Protein
-
Green Fluorescent Proteins
-
beta-Galactosidase