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Published online 16 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 7 2123-2128
© 2004 Oxford University Press

The RNA-dependent RNA polymerase, QDE-1, is a rate-limiting factor in post-transcriptional gene silencing in Neurospora crassa

Emma C. Forrest, Carlo Cogoni and Giuseppe Macino*

Istituto Pasteur e Fondazione Cenci Bolognetti, Dipartimento di Biotecnologie Cellulari ed Ematologia, Sezione di Genetica Molecolare, Universitá di Roma La Sapienza, 00161 Roma, Italy

*To whom correspondence should be addressed. Tel: +39 064452806; Fax: +39 064457731; Email: macino{at}bce.uniroma1.it

Received February 12, 2004; Revised and Accepted March 17, 2004

The RNA-dependent RNA polymerase (RdRP) qde-1 is an essential component of post-transcriptional gene silencing (PTGS), termed ‘quelling’ in the fungus Neurospora crassa. Here we show that the overexpression of QDE-1 results in a dramatic increase in the efficiency of quelling, with a concomitant net increase in the quantity of al-1 siRNAs. Moreover, in overexpressed strains there is a significant reduction in the number of transgenes required to induce quelling, and an increase in the phenotypic stability despite progressive loss of tandemly repeated transgenes, which normally determines reversion of a silenced phenotype to wild type. These data suggest that the activation and maintenance of silencing in Neurospora appear to rely both on the cellular amount of QDE-1 and the amount of transgenic copies producing RNA molecules that act as a substrate for the RdRP, implicating QDE-1 as a rate-limiting factor in PTGS.


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