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PMID:15355351

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Citation

Tetsuka, T, Uranishi, H, Sanda, T, Asamitsu, K, Yang, JP, Wong-Staal, F and Okamoto, T (2004) RNA helicase A interacts with nuclear factor kappaB p65 and functions as a transcriptional coactivator. Eur. J. Biochem. 271:3741-51

Abstract

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 kappaB (NF-kappaB). The interaction between p65 and RHA was confirmed by glutathione-S transferase pull-down assay in vitro, and by co-immunoprecipitation assay in vivo. In transient transfection assays, RHA enhanced NF-kappaB dependent reporter gene expression induced by p65, tumor necrosis factor-alpha, or NF-kappaB inducing kinase. The mutant form of RHA lacking ATP-binding activity inhibited NF-kappaB dependent reporter gene expression induced by these activators. Moreover, depletion of RHA using short interfering RNA reduced the NF-kappaB dependent transactivation. These data suggest that RHA is an essential component of the transactivation complex by mediating the transcriptional activity of NF-kappaB.

Links

PubMed Online version:10.1111/j.1432-1033.2004.04314.x

Keywords

Autoantigens/metabolism; Blotting, Western; Catalysis; Cell Line; DEAD-box RNA Helicases; Gene Expression; Genes, Reporter; Humans; NF-kappa B/chemistry; NF-kappa B/metabolism; Neoplasm Proteins; Precipitin Tests; Protein Binding; Protein Structure, Tertiary; Protein Subunits/metabolism; RNA Helicases/metabolism; RNA Interference; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/metabolism; Saccharomyces cerevisiae/genetics; Trans-Activators/metabolism; Transcription, Genetic; Transcriptional Activation; Two-Hybrid System Techniques

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:TF65

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q08211

F

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q04206

F

Seeded From UniProt

complete

HUMAN:DHX9

involved_in

GO:0051092: positive regulation of NF-kappaB transcription factor activity

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DHX9

involved_in

GO:0071356: cellular response to tumor necrosis factor

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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