RNA干扰与miRNAPPT.ppt

Fig. 2. Structure and function of Arabidopsis miRNAs. DCL1 expression (it catalyzes pre-miRNA processing, is under negative-feedback regulation by miR162 (left). miR165/166 negatively regulates PHV and PHB mRNAs by guiding sequence-specific cleavage (right). PHV and PHB both encode HD-Zip transcription factors. miR165 and miR166 are related miRNAs that likely interact with PHV and PHB mRNAs (PHV mRNA and miR165 shown). Arrow, miR165-guided cleavage site. (B) Model for specification of adaxial/abaxial polarity in Arabidopsis leaves. Expression of PHV/PHB in leaf primordium cells close to the meristem results in the adaxial transcriptional program. Inhibition of PHV/PHB by miR165/166-guided degradation in cells distal to the meristem specifies abaxial fate. Negative feedback loops in miRNA processing Ago family - phylogenetic distribution General: Mammalian Ago family identified a few years ago. Slicer function of mammalian Ago2 well understood. But why is only human Ago2 a Slicer endonuclease, when the catalytic residues are conserved in other Ago proteins? How might the modification of Ago proteins (e.g., phosphorylation) affect Ago activity? What are the cellular partners of Agos, and how do they affect/contribute to their function? Ago function: remove passenger strand from dsRNA, for guide strand to enter mature miRNP [Cell (2007), Diederichs Haber] During microRNA biogenesis, Drosha Dicer process pri-miRNA through premiRNA hairpins to mature miRNA, which assembels into the RNA-Induced Silencing Complex (RISC) with Ago proteins, the effector molecules for RNAi. This paper: all Argonautes elevate mature miRNA expression post-transcriptionally, independent of RNase activity. Also, Ago2 cleaves the pre-miRNA to an additional processing intermediate, termed Ago2-cleaved precursor miRNA or ac-pre-miRNA. This on-pathway intermediate cleaves 12 nt from pre-miRNA 3’ end. By analogy to siRNA, Ago2 cleavage facilitates