GH_3-26-03.ppt

miRNAs and other RNAi-related factors: functions in plant and animal gene regulation and development Products of dsRNA cleavage regulate gene expression exogenous dsRNA triggers RNA interference RNAi serves as viral/transposon defense mechanism endogenous mRNAs may be cleaved by an RNAi-like mechanism siRNAs may guide promoter methylation (TGS) miRNAs produced from a hairpin precursor inhibit translation (among other effects?) Mechanism of RNAi heterochronic mutations cause lineage defects sin1/caf mutations delay flowering and increase leaf number Mutations in sin1 alter ovule development SIN1 mRNA is expressed in floral tissue; sin1 affects ovules Expression: meristems, flower, early ovule/embryo, stigma, vasculature GUS reporter (partly) recapitulates in situ Transcriptionally active SIN1 allele is supplied maternally sin1/caf mutations affect floral architecture sin1 mutations are expected to affect RNA binding or unwinding similar molecules in Arabidopsis and other critters SIN1/CAF might processs dsRNA and/or act in miRNA metabolism Transgene overexpression can trigger silencing of the transgene and the endogenous locus The silencing signal spreads through the vasculature Model of silencing and viral suppressor action HC-Pro expression suppresses growth HC-Pro expression causes floral developmental defects HC-Pro expression and dcl1 mutations cause similar phenotypes HC-Pro and dcl1 interfere with mRNA cleavage wT HC-Pro inhibits processing of coexpressed SCL6-IV mRNA miRNA levels increased by HC-Pro, decreased by dcl1 mRNAs are cleaved within mRNA-miRNA duplexed region HC-Pro expression blocks cleavage/degradation of mRNAs HC-Pro inhibits accumulation of siRNAs Future directions How does CAF/DCL1 function in early development? Which DCL1 paralog functions in: production of siRNAs? cleavage of mRNAs targeted by miRNAs? What are the targets of plant miRNAs? Do any function in translational inhibition? Any function in transcriptional silencing? How do miRNA