An in vitro enzymatic assay coupled to proteomics analysis reveals a new DNA processing activity for Ewing sarcoma and TAF(II)68 proteins.
Based on structural and functional similarities, translocated in liposarcoma/fusion (TLS/FUS) protein, Ewing sarcoma (EWS) protein and human TATA binding protein-associated factor (hTAF(II)68) have been grouped in the TLS-EWS-TAF(II)68 (TET) protein family. Translocations involving their genes lead to sarcomas. Polypyrimidine tract-binding protein-associated splicing factor (PSF), although not grouped in this family, presents structural and functional similarities with TET proteins and is involved in translocation leading to carcinoma. Beside their role in RNA metabolism, the precise cellular functions of these multifunctional proteins are not yet fully elucidated. We previously showed that both TLS/FUS and PSF display activities able to pair homologous DNA on membrane in an in vitro assay. In the present study, we address the question whether EWS and hTAF(II)68 also display pairing on membrane activities, and to a larger extent whether other proteins also exhibit such activity. We applied the pairing on membrane assay to 2-DE coupled to MS analysis for a global screening of DNA pairing on membrane activities. In addition to TLS/FUS and PSF, this test allowed us to identify EWS and hTAF(II)68, but no other proteins, indicating a feature specific to a protein family whose members share extensive structural similarities. This common activity suggests a role for TET proteins and PSF in genome plasticity control.
Guipaud
Olivier
O
Commissariat à l'Energie Atomique, Département de Radiobiologie et Radiopathologie, Fontenay aux Roses, France.
Guillonneau
François
F
Labas
Valérie
V
Praseuth
Danièle
D
Rossier
Jean
J
Lopez
Bernard
B
Bertrand
Pascale
P
D002461:Cell Line, Transformed; D004273:DNA, Neoplasm / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D010766:Phosphorylation; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D034802:RNA-Binding Protein EWS / Q000378:metabolism*; D034702:RNA-Binding Protein FUS / Q000378:metabolism; D035381:TATA-Binding Protein Associated Factors / Q000378:metabolism*
D004273:DNA, Neoplasm; D020543:Proteome; D034802:RNA-Binding Protein EWS; D034702:RNA-Binding Protein FUS; D035381:TATA-Binding Protein Associated Factors