Intracellular characterization of DDX39, a novel growth-associated RNA helicase

DDX39 belongs to the DEAD box RNA helicase family and is overexpressed in human lung squamous cell carcinoma. In this study, in order to seek the biological relevance of DDX39, we conducted its intracellular characterization. When expressed in 293 cells, DDX39 undergoes heavy ubiquitylation and the stability of DDX39 is regulated via a ubiqutin-proteasome pathway. DDX39 tethers ALY, an essential mRNA export factor, in vivo, confirming the role of DDX39 in the RNA splicing/export process. Co-immunoprecipitation and mass spectrometry analyses detected CIP29, a recently discovered growth and cell cycle-related factor, as a main DDX39-interacting protein. CIP29 binds RNA on its own and enhances RNA unwinding activity of DDX39. Thus, CIP29 physically and functionally associates with DDX39, suggesting their cooperation in the RNA metabolism. Extension of the search for the protein-protein interactions encompassing DDX39 identified FUS/TLS, a nucleic acid binding protein participating in both transcription and splicing, as a CIP29-interacting protein. The connections comprising ALY, DDX39, CIP29 and FUS/TLS may be an integral part of transcription, splicing and RNA export. We simultaneously examined the properties of DDX39-S, a C-terminally truncated variant of DDX39 stemmed from alternative splicing, to understand its biological significance.