(精编)molecular-biology-2010_autumn-15.pptVIP

Post-Transcriptional Regulation of Eukaryotic Genes (真核基因的转录后调控) RNA silencing (siRNA and miRNA) Protein degradation;Gene Silencing;DNA; Terminology ;1. RNAi的发现;Double-stranded RNA is the trick!!!!!;小RNA的发现;David Baulcombe;;?The PAZ domain binds the 2?nt 3′ overhang of a dsRNA terminus. The RNaseIII domains form a pseudo-dimer. Each domain hydrolyzes one strand of the substrate. The binding site of the dsRBD is not defined. Hammond (2005) FEBS Lett 579:5822-5829 ;A model for dsRNA processing by Dicer;siRNA 的装载与激活（Loading activation of siRNA ）;siRNA asymmetry;; The PAZ domain PIWI is an RNase H domain;The siRNA guide strand is bound at the 5′end by the PIWI domain and at the 3′end by the PAZ domain. mRNA targets are initially bound by the seed region of the siRNA and pairing is extended to the 3′end. Slicer cleavage is measured from the 5′end of the siRNA. ;RNA依赖的RNA聚合酶（RNA-dependent RNA polymerase ，RDRP);a, RNAs are normally not silenced because the RDR proteins do not have access to the template RNA sequence. Cap-binding protein (CBP) and poly-adenosine-binding protein (PABP) may be involved in this restriction of RDR access. However in b, the RDR protein is allowed access because the RNA lacks a 5 cap or 3 poly-adenosine tail, and dsRNA is produced which enters the siRNA pathway. b, The amplification process would result from the ability of a single aberrant RNA to generate many molecules of siRNA. c shows the outcome if a small quantity of primary siRNA is present from either a virus, a transposon or from a cellular RNA through the process shown in b. The antisense strand of this siRNA may anneal by base pairing to a target RNA and serve as a primer for the RDR. The resulting dsRNA would then be cleaved by Dicer and, as in b, there would be amplification because many secondary siRNAs would be produced from each molecule of primary siRNA.;RNA dependent DNA methylation (RdDM);;Short and long range cell to cell movement of RNAi;;RNAi的生物学意义;microRN