title: Diverse gene-silencing mechanisms with distinct requirements for RNA polymerase subunits in Zea mays.
abstract: In Zea mays, transcriptional regulation of the b1 (booster1) gene requires a distal enhancer and MEDIATOR OF PARAMUTATION1 (MOP1), MOP2, and MOP3 proteins orthologous to Arabidopsis components of the RNA-dependent DNA methylation pathway. We compared the genetic requirements for MOP1, MOP2, and MOP3 for endogenous gene silencing by two hairpin transgenes with inverted repeats of the a1 (anthocyaninless1) gene promoter (a1pIR) and the b1 gene enhancer (b1IR), respectively. The a1pIR transgene induced silencing of endogenous A1 in mop1-1 and mop3-1, but not in Mop2-1 homozygous plants. This finding suggests that transgene-derived small interfering RNAs (siRNAs) circumvented the requirement for MOP1, a predicted RNA-dependent RNA polymerase, and MOP3, the predicted largest subunit of RNA polymerase IV (Pol IV). Because the Arabidopsis protein orthologous to MOP2 is the second largest subunit of Pol IV and V, our results may indicate that hairpin-induced siRNAs cannot bypass the requirement for the predicted scaffolding activity of Pol V. In contrast to a1pIR, the b1IR transgene silenced endogenous B1 in all three homozygous mutant genotypes--mop1-1, Mop2-1, and mop3-1--suggesting that transgene mediated b1 silencing did not involve MOP2-containing Pol V complexes. Based on the combined results for a1, b1, and three previously described loci, we propose a speculative hypothesis of locus-specific deployment of Pol II, MOP2-containing Pol V, or alternative versions of Pol V with second largest subunits other than MOP2 to explain the mechanistic differences in silencing at specific loci, including one example associated with paramutation.
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, Florida 32306-4295.
authors.forename: Amy E
authors.initials: AE
authors.lastname: Sloan
authors.affiliation: BIO5 Institute and Department of Plant Science, University of Arizona, Tucson, Arizona 85721.
authors.forename: Lyudmila
authors.initials: L
authors.lastname: Sidorenko
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, Florida 32306-4295 mcginnis@bio.fsu.edu.
authors.forename: Karen M
authors.initials: KM
authors.lastname: McGinnis
mesh_terms: D020090:Chromosome Segregation / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics; D020868:Gene Silencing; D017343:Genes, Plant; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D010860:Pigments, Biological / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D021122:Protein Subunits / Q000235:genetics* / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D019076:Transgenes; D003313:Zea mays / Q000201:enzymology* / Q000235:genetics*
chemical_list: D010860:Pigments, Biological; D010940:Plant Proteins; D021122:Protein Subunits; D012321:DNA-Directed RNA Polymerases
keywords: RNA polymerase IV/V; RNA-directed DNA methylation; epigenetics; paramutation; transcriptional gene silencing