Abstract
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.
MeSH Terms
Fungal Proteins/genetics,metabolism
Gene Expression Regulation, Fungal
Gene Silencing
Neurospora crassa/enzymology,genetics
Phenotype
RNA, Fungal/genetics,metabolism
RNA, Messenger/genetics,metabolism
RNA-Dependent RNA Polymerase/genetics,metabolism
Transcription, Genetic
Transformation, Genetic
Chemicals
Fungal Proteins
RNA, Fungal
RNA, Messenger
QDE1 protein, Neurospora crassa
RNA-Dependent RNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Forrest Emma C
Istituto Pasteur e Fondazione Cenci Bolognetti, Dipartimento di Biotecnologie Cellulari ed Ematologia, Sezione di Genetica Molecolare, Universitá di Roma La Sapienza, 00161 Roma, Italy.
Cogoni Carlo
Macino Giuseppe
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