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Papers In Press, published online ahead of print October 30, 2001
Biochemistry, Institute of Molecular Biotechnology, Jena D-07745
Corresponding Author: fgrosse{at}imb-jena.de
Nuclear DNA helicase II (NDH II), also designated RNA helicase A (RHA), is a multifunctional protein involved in transcription, RNA processing and transport. Here we report that NDH II binds to filamentous actin (F-actin). NDH II was partially purified from HeLa nuclear extract by ion-exhange chromatography on Bio-Rex 70 and DEAE-Sepharose. Upon gel filtration chromatography on Sepharose 4B partially purified NDH II resolved into two distinct peaks. The first NDH II peak, corresponding to the void volume of Sepharose 4B, displayed co-elution with an abundant 42 kDa protein that was subsequently identified as actin. Several nuclear proteins such as RNA polymerase II, the U5 snRNP-associated WD40 protein and hnRNP proteins co-purified with NDH II. However, only hnRNP A1 and C were found together with NDH II and actin polymers during gel filtration. NDH II and hnRNP C from the HeLa nuclear extract co-eluted with F-actin on Sepharose 4B in an RNase-resistant manner, while hnRNP A1 was nearly completely removed from F-actin-associated hnRNP complexes following RNA digestion. The association of NDH II and hnRNP C with F-actin was abolished by gelsolin, an F-actin depolymerizing protein that fragments actin polymers into oligomers or monomers. Furthermore, NDH II co-immunoprecipitated with F-actin and hnRNP C, respectively. In vitro translated NDH II co-eluted with F-actin on Sepharose 4B, whereas no co-elution with F-actin was observed for the in vitro translated hnRNP A1 or hnRNP C1. Binding to F-actin requires an intact C-terminus of NDH II and most likely a native protein conformation. Electron microscopy indicated a close spatial proximity between NDH II, hnRNP C and F-actin within the HeLa nucleus. These results suggest an important function of NDH II in mediating the attachment of hnRNP/mRNP complexes to the actin nucleoskeleton for RNA processing, transport, or other actin-related processes.
J. Biol. Chem, 10.1074/jbc.M109393200
Submitted on September 28, 2001
Revised on October 24, 2001
Accepted on October 29, 2001
Nuclear DNA helicase II/RNA helicase A binds to filamentous actin (F-actin)
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