(精选)molecular-biology-2010_autumn-15教学课件.pptVIP

The PAZ domain PIWI is an RNase H domain A binding pocket in PAZ accommodates the 2 nt overhang No interactions found between the 2 nts with the pocket, suggesting that the pocket accommodates all nucleotide combinations Ma et al. Nature 429, 318-322 (2004) The similarity was not obvious at the primary sequence level !! Song et al. Science 305, 1434-1437. 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. Model for Slicer catalysis RNA依赖的RNA聚合酶（RNA-dependent RNA polymerase ，RDRP) 最早克隆自被类病毒侵染的番茄中 (1998, Plant Cell) 与RNA病毒编码的RdRP 基本没有同源性 在果蝇和哺乳动物基因组中不存在 是产生次级小干扰RNA的放大效应的主要因子 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 RNA polymerase (RDR) transitive RNAi RNA dependent DNA methylation (RdDM) RdDM主要通过dsRNA介导相同DNA序列发生重新甲基化(de novo methylation)而实现转录水平的基因沉默。 引起相同序列DNA甲基化的物质是dsRNA RdDM 的生物学意义: 阻抑不必要基因（repetitive sequences)和有害基因(尤其