Identification of hnRNP P2 as TLS/FUS using electrospray mass spectrometry.
Protein complexes assembled on mRNA precursors can be separated by gel filtration chromatography to yield spliceosomal and H complex fractions (Reed R, Griffith J, Maniatis T, 1988, Cell 53:949-961; Reed R, 1990, Proc Natl Acad Sci USA 87:8031-8035.). Here we use Nano electrospray mass spectrometry (Wilm M, Mann M, 1994, Int J Mass Spectrometry Ion Processes 136:167-180) to identify proteins complexed with Adeno-pre-mRNA in the H complex peak. Four of the major hnRNP proteins, A1, B1, C1, and G, were identified by database analysis based on peptide mass and sequence information. A fifth protein in the H complex peak, corresponding to hnRNP P2, is shown to be the product of the TLS/FUS gene. This was originally identified as a chimeric oncogene formed by the chromosome translocation t(12;16) that is responsible for myxoid liposarcoma. The involvement of hnRNP P2 in oncogenesis provides a clear example of the importance of hnRNP proteins in molecular disease.
Calvio
C
C
Gene Expression Programme, European Molecular Biology Laboratory, Heidelberg.
Neubauer
G
G
Mann
M
M
Lamond
A I
AI
D000595:Amino Acid Sequence; D001483:Base Sequence; D002467:Cell Nucleus / Q000737:chemistry; D006367:HeLa Cells; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry*; D009687:Nuclear Proteins / Q000737:chemistry*; D034702:RNA-Binding Protein FUS; D016601:RNA-Binding Proteins / Q000737:chemistry*; D012261:Ribonucleoproteins / Q000737:chemistry*; D017421:Sequence Analysis
C439788:C1 HNRNP; C503635:HNRNPC protein, human; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D034702:RNA-Binding Protein FUS; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C439768:hnRNP A2