Proto-oncoprotein TLS/FUS is associated to the nuclear matrix and complexed with splicing factors PTB, SRm160, and SR proteins.
TLS/FUS is a nucleic acid-binding protein whose N-terminal half functions as a transcriptional activator domain in fusion oncoproteins found in human leukemias and liposarcomas. Previous reports have suggested a role for TLS/FUS in transcription and splicing processes. Here we report the association of TLS/FUS with the nuclear matrix and investigate ... its role in splicing. Splicing of two pre-mRNAs was inhibited in a TLS/FUS-immunodepleted extract and could only be partly restored by addition of recombinant TLS/FUS or/and SR proteins, known interaction partners of TLS/FUS. The subsequent analysis of TLS/FUS immunoprecipitates revealed that, in addition to the SR proteins SC35 and SRp75, the splicing factor PTB (hnRNPI) and the splicing coactivator SRm160 are complexed with TLS/FUS, thus explaining the inability to restore splicing completely. Coimmunolocalization confirmed the nuclear matrix association and interaction of TLS/FUS with PTB, SR proteins, and SRm160. Our results suggest that the matrix protein TLS/FUS plays a role in spliceosome assembly.
Mesh Terms:
Antigens, Nuclear, Hela Cells, Humans, Microscopy, Fluorescence, Nuclear Matrix, Nuclear Matrix-Associated Proteins, Polypyrimidine Tract-Binding Protein, Precipitin Tests, Protein Binding, Proto-Oncogene Proteins, RNA Splicing, RNA-Binding Protein FUS, RNA-Binding Proteins, Signal Recognition Particle, Spliceosomes
Antigens, Nuclear, Hela Cells, Humans, Microscopy, Fluorescence, Nuclear Matrix, Nuclear Matrix-Associated Proteins, Polypyrimidine Tract-Binding Protein, Precipitin Tests, Protein Binding, Proto-Oncogene Proteins, RNA Splicing, RNA-Binding Protein FUS, RNA-Binding Proteins, Signal Recognition Particle, Spliceosomes
Exp. Cell Res.
Date: Feb. 15, 2003
PubMed ID: 12581738
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