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Nucleic Acids Research 2006 34(3):773-784; doi:10.1093/nar/gkj465
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Published online 2 February 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
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Article

HelF, a putative RNA helicase acts as a nuclear suppressor of RNAi but not antisense mediated gene silencing

Blagovesta Popova, Markus Kuhlmann, Andrea Hinas1, Fredrik Söderbom1 and Wolfgang Nellen*

Abt. Genetik, Universität Kassel Heinrich-Plett-Strasse 40, D-34132 Kassel, Germany 1Department of Molecular Biology, Biomedical Center, Swedish University of Agricultural Sciences Box 590, S-75124 Uppsala, Sweden

*To whom correspondence should be addressed. Tel: +49 0 561 804 4805; Fax: +49 0 561 804 4800; Email: nellen{at}uni-kassel.de

Received December 12, 2005. Revised January 9, 2006. Accepted January 9, 2006.

We have identified a putative RNA helicase from Dictyostelium that is closely related to drh-1, the ‘dicer-related-helicase’ from Caenorhabditis elegans and that also has significant similarity to proteins from vertebrates and plants. Green fluorescent protein (GFP)-tagged HelF protein was localized in speckles in the nucleus. Disruption of the helF gene resulted in a mutant morphology in late development. When transformed with RNAi constructs, HelF– cells displayed enhanced RNA interference on four tested genes. One gene that could not be knocked-down in the wild-type background was efficiently silenced in the mutant. Furthermore, the efficiency of silencing in the wild-type was dramatically improved when helF was disrupted in a secondary transformation. Silencing efficiency depended on transcription levels of hairpin RNA and the threshold was dramatically reduced in HelF– cells. However, the amount of siRNA did not depend on hairpin transcription. HelF is thus a natural nuclear suppressor of RNA interference. In contrast, no improvement of gene silencing was observed when mutant cells were challenged with corresponding antisense constructs. This indicates that RNAi and antisense have distinct requirements even though they may share parts of their pathways.

DDBJ/EMBL/GenBank accession no. HelF: DDB0168963


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