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Evidence that RNA silencing functions as an antiviral defense mechanism in fungi.

ABSTRACT: The role of RNA silencing as an antiviral defense mechanism in fungi was examined by testing the effect of dicer gene disruptions on mycovirus infection of the chestnut blight fungus Cryphonectria parasitica. C. parasitica dicer-like genes dcl-1 and dcl-2 were cloned and shown to share a high level of predicted amino acid sequence identity with the corresponding dicer-like genes from Neurospora crassa [Ncdcl-1 (50.5%); Ncdcl-2 (38.0%)] and Magnaporthe oryzae [MDL-1 (45.6%); MDL-2 (38.0%)], respectively. Disruption of dcl-1 and dcl-2 resulted in no observable phenotypic changes relative to wild-type C. parasitica. Infection of Deltadcl-1 strains with hypovirus CHV1-EP713 or reovirus MyRV1-Cp9B21 resulted in phenotypic changes that were indistinguishable from that exhibited by wild-type strain C. parasitica EP155 infected with these same viruses. In stark contrast, the Deltadcl-2 and Deltadcl-1/Deltadcl-2 mutant strains were highly susceptible to mycovirus infection, with CHV1-EP713-infected mutant strains becoming severely debilitated. Increased viral RNA levels were observed in the Deltadcl-2 mutant strains for a hypovirus CHV1-EP713 mutant lacking the suppressor of RNA silencing p29 and for wild-type reovirus MyRV1-Cp9B21. Complementation of the Deltadcl-2 strain with the wild-type dcl-2 gene resulted in reversion to the wild-type response to virus infection. These results provide direct evidence that a fungal dicer-like gene functions to regulate virus infection.

SUBMITTER: Segers GC

PROVIDER: S-EPMC1937564 | biostudies-literature | 2007 Jul

REPOSITORIES: biostudies-literature

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Evidence that RNA silencing functions as an antiviral defense mechanism in fungi.

Segers Gert C GC Zhang Xuemin X Deng Fuyou F Sun Qihong Q Nuss Donald L DL

Proceedings of the National Academy of Sciences of the United States of America 20070723 31

The role of RNA silencing as an antiviral defense mechanism in fungi was examined by testing the effect of dicer gene disruptions on mycovirus infection of the chestnut blight fungus Cryphonectria parasitica. C. parasitica dicer-like genes dcl-1 and dcl-2 were cloned and shown to share a high level of predicted amino acid sequence identity with the corresponding dicer-like genes from Neurospora crassa [Ncdcl-1 (50.5%); Ncdcl-2 (38.0%)] and Magnaporthe oryzae [MDL-1 (45.6%); MDL-2 (38.0%)], respe ...[more]

PMID: 17646660

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Project description:The increasing knowledge on the functional relevance of endogenous small RNAs (esRNAs) as riboregulators has stimulated the identification and characterization of these molecules in numerous eukaryotes. In the basal fungus Mucor circinelloides, an emerging opportunistic human pathogen, esRNAs that regulate the expression of many protein coding genes have been described. These esRNAs share common machinery for their biogenesis consisting of an RNase III endonuclease Dicer, a single Argonaute protein and two RNA-dependent RNA polymerases. We show in this study that, besides participating in this canonical dicer-dependent RNA interference (RNAi) pathway, the rdrp genes are involved in a novel dicer-independent degradation process of endogenous mRNAs. The analysis of esRNAs accumulated in wild type and silencing mutants demonstrates that this new rdrp-dependent dicer-independent regulatory pathway, which does not produce sRNA molecules of discrete sizes, controls the expression of target genes promoting the specific degradation of mRNAs by a previously unknown RNase. This pathway mainly regulates conserved genes involved in metabolism and cellular processes and signaling, such as those required for heme biosynthesis, and controls responses to specific environmental signals. Searching the Mucor genome for candidate RNases to participate in this pathway, and functional analysis of the corresponding knockout mutants, identified a new protein, R3B2. This RNase III-like protein presents unique domain architecture, it is specifically found in basal fungi and, besides its relevant role in the rdrp-dependent dicer-independent pathway, it is also involved in the canonical dicer-dependent RNAi pathway, highlighting its crucial role in the biogenesis and function of regulatory esRNAs. The involvement of RdRPs in RNA degradation could represent the first evolutionary step towards the development of an RNAi mechanism and constitutes a genetic link between mRNA degradation and post-transcriptional gene silencing.

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Evidence that RNA silencing functions as an antiviral defense mechanism in fungi.

Publications

Evidence that RNA silencing functions as an antiviral defense mechanism in fungi.

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