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Molecular and Cellular Biology, March 2004, p. 2536-2545, Vol. 24, No. 6
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.6.2536-2545.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Redundancy of the Two Dicer Genes in Transgene-Induced Posttranscriptional Gene Silencing in Neurospora crassa{dagger}

Caterina Catalanotto,1 Massimiliano Pallotta,1 Paul ReFalo,2 Matthew S. Sachs,2,3 Laurence Vayssie,1 Giuseppe Macino,1 and Carlo Cogoni1*

Dipartimento di Biotecnologie Cellulari ed Ematologia, Sezione di Genetica Molecolare, Universita' di Roma La Sapienza, 00161 Rome, Italy,1 Department of Environmental and Biomolecular Systems, OGI School of Science and Engineering, Oregon Health and Science University, Beaverton, Oregon 97006,2 Department of Molecular Microbiology and Immunology, School of Medicine, Oregon Health and Science University, Portland, Oregon 972013

Received 31 July 2003/ Returned for modification 17 November 2003/ Accepted 16 December 2003

RNA interference (RNAi) in animals, cosuppression in plants, and quelling in fungi are homology-dependent gene silencing mechanisms in which the introduction of either double-stranded RNA (dsRNA) or transgenes induces sequence-specific mRNA degradation. These phenomena share a common genetic and mechanistic basis. The accumulation of short interfering RNA (siRNA) molecules that guide sequence-specific mRNA degradation is a common feature in both silencing mechanisms, as is the component of the RNase complex involved in mRNA cleavage. During RNAi in animal cells, dsRNA is processed into siRNA by an RNase III enzyme called Dicer. Here we show that elimination of the activity of two Dicer-like genes by mutation in the fungus Neurospora crassa eliminates transgene-induced gene silencing (quelling) and the processing of dsRNA to an siRNA form. The two Dicer-like genes appear redundant because single mutants are quelling proficient. This first demonstration of the involvement of Dicer in gene silencing induced by transgenes supports a model by which a dsRNA produced by the activity of cellular RNA-dependent RNA polymerases on transgenic transcripts is an essential intermediate of silencing.

* Corresponding author. Mailing address: Dipartimento di Biotecnologie Cellulari ed Ematologia, Sezione di Genetica Molecolare, Universita' di Roma La Sapienza, Viale Regina Elena 324, 00161 Rome, Italy. Phone: 39 06 4457731. Fax: 39 06 4457731. E-mail: carlo{at}bce.uniroma1.it.

{dagger} This work is dedicated to the memory of our colleague Giusi Arpaia.

Molecular and Cellular Biology, March 2004, p. 2536-2545, Vol. 24, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.6.2536-2545.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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