TAILIEUCHUNG - Báo cáo khoa học: RNA helicase A interacts with nuclear factor jB p65 and functions as a transcriptional coactivator

RNA helicase A (RHA), a member of DNA and RNA helicase family containing ATPase activity, is involved in many steps of gene expression such as transcription and mRNA export. RHA has been reported to bind directly to the transcriptional coactivator, CREB-binding protein, and the tumor suppressor protein, BRCA1, and links them to RNAPolymerase II holoenzyme complex. Using yeast two-hybrid screening, we have identified RHA as an interacting molecule of the p65 subunit of nuclear factorjB(NF-jB). | Eur. J. Biochem. 271 3741-3751 2004 FEBS 2004 doi RNA helicase A interacts with nuclear factor kB p65 and functions as a transcriptional coactivator Toshifumi Tetsuka1 Hiroaki Uranishi1 Takaomi Sanda1 Kaori Asamitsu1 Jiang-Ping Yang2 Flossie Wong-Staal2 and Takashi Okamoto1 1 Department of Molecular and Cellular Biology Nagoya City University Graduate School of Medical Sciences Nagoya Aichi Japan department of Medicine University of California San Diego La Jolla CA USA RNA helicase A RHA a member of DNA and RNA helicase family containing ATPase activity is involved in many steps of gene expression such as transcription and mRNA export. RHA has been reported to bind directly to the transcriptional coactivator CREB-binding protein and the tumor suppressor protein BRCA1 and links them to RNA Polymerase II holoenzyme complex. Using yeast two-hybrid screening we have identified RHA as an interacting molecule of the p65 subunit of nuclear factor kB NF-kB . The interaction between p65 and RHA was confirmed by glutathione-S transferase pull-down assay in vitro and by coimmunoprecipitation assay in vivo. In transient transfection assays RHA enhanced NF-kB dependent reporter gene expression induced by p65 tumor necrosis factor-a or NF-kB inducing kinase. The mutant form of RHA lacking ATP-binding activity inhibited NF-kB dependent reporter gene expression induced by these activators. Moreover depletion of RHA using short interfering RNA reduced the NF-kB dependent transactivation. These data suggest that RHA is an essential component of the transactivation complex by mediating the transcriptional activity of NF-kB. Keywords coactivator NF-kB protein-protein interaction RNA helicase A transcription. Nuclear factor kB NF-kB is an inducible cellular transcription factor that regulates a wide variety of cellular and viral genes including cytokines cell adhesion molecules and HIV 1-3 . The members of the NF-kB family in mammalian cells include the

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