Interaction of RNA polymerase II and the small RNA machinery affects heterochromatic silencing in Drosophila.
Interaction of RNA polymerase II and the small RNA machinery affects heterochromatic silencing in Drosophila.
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RNA聚合酶II和小的RNA机械的相互作用会影响果蝇中的异质沉默。
DOI：
10.1186/1756-8935-2-15
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发表时间：
2009-11-16
影响因子：
3.9
通讯作者：
Birchler JA
中科院分区：
文献类型：
--
作者：
Kavi HH;Birchler JA
异染色质是真核生物染色体上紧密包裹的动态区域，在有丝分裂和减数分裂重组等细胞过程中起着至关重要的作用。最近对裂糖菌的实验表明，着丝粒异染色质的结构在RNAi途径突变体中受到影响。研究还表明，在裂变酵母中，RNA Pol II穿过异染色质屏障，RNA Pol II通过异染色质对异染色质结构很重要。因此，RNAi机制和RNA Pol II之间复杂的相互作用影响了异染色质结构。然而，RNAi机制和RNA Pol II在后生动物异染色质景观中的作用尚不清楚。本研究分析了小RNA机制和RNA Pol II在果蝇异染色质结构中的相互作用。研究结果表明，RNA Pol II（最大和第二大亚基）与RNA沉默机制小组分（dcr-2、ago1、ago2、piwi、Lip [D]、aub和hls）之间存在遗传和生化相互作用。对RNA Pol II反杂合子多烯染色体和不同小RNA通路突变的免疫荧光分析显示H3K9me2减少和异染色质蛋白-1错位。对这些突变体进行的遗传分析表明，这些突变体对白斑4h位置效应有很强的抑制作用。western blot分析和染色质免疫沉淀进一步证实了这一点，结果显示，与野生型或单杂合子相比，反杂合子突变体的H3K9me2减少。用果蝇胚胎提取物进行共免疫沉淀，发现RNA Pol II最大亚基与Dcr-2和dAGO1相互作用。在多粒染色体上进行的共定位显示RNA Pol II和dAGO1在某些位点重叠。我们的实验显示了果蝇RNA Pol II（最大和第二大亚基）和小RNA沉默机制之间的遗传和生化相互作用。相互作用在确定H3K9me2和HP-1沉积在色中心异染色质方面具有功能方面。因此，RNA Pol II在果蝇异染色质结构的建立中具有重要作用。
Heterochromatin is the tightly packaged dynamic region of the eukaryotic chromosome that plays a vital role in cellular processes such as mitosis and meiotic recombination. Recent experiments in Schizosaccharomyces pombe have revealed the structure of centromeric heterochromatin is affected in RNAi pathway mutants. It has also been shown in fission yeast that the heterochromatin barrier is traversed by RNA Pol II and that the passage of RNA Pol II through heterochromatin is important for heterochromatin structure. Thus, an intricate interaction between the RNAi machinery and RNA Pol II affects heterochromatin structure. However, the role of the RNAi machinery and RNA Pol II on the metazoan heterochromatin landscape is not known. This study analyses the interaction of the small RNA machinery and RNA Pol II on Drosophila heterochromatin structure. The results in this paper show genetic and biochemical interaction between RNA Pol II (largest and second largest subunit) and small RNA silencing machinery components (dcr-2, ago1, ago2, piwi, Lip [D], aub and hls). Immunofluorescence analysis of polytene chromosomes from trans-heterozygotes of RNA Pol II and different mutations of the small RNA pathways show decreased H3K9me2 and mislocalization of Heterochromatin protein-1. A genetic analysis performed on these mutants showed a strong suppression of white-mottled4h position effect variegation. This was further corroborated by a western blot analysis and chromatin immunoprecipitation, which showed decreased H3K9me2 in trans-heterozygote mutants compared to wild type or single heterozygotes. Co-immunoprecipitation performed using Drosophila embryo extracts showed the RNA Pol II largest subunit interacting with Dcr-2 and dAGO1. Co-localization performed on polytene chromosomes showed RNA Pol II and dAGO1 overlapping at some sites. Our experiments show a genetic and biochemical interaction between RNA Pol II (largest and second largest subunits) and the small RNA silencing machinery in Drosophila. The interaction has functional aspects in terms of determining H3K9me2 and HP-1 deposition at the chromocentric heterochromatin. Thus, RNA Pol II has an important role in establishing heterochromatin structure in Drosophila.
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影响因子：
2.7
作者：
Chanas, G;Lavrov, S;Maschat, F
通讯作者：
Maschat, F
影响因子：
30.8
作者：
Noma, K;Sugiyama, T;Grewal, SIS
通讯作者：
Grewal, SIS
影响因子：
64.5
作者：
Jia, ST;Yamada, T;Grewal, SIS
通讯作者：
Grewal, SIS
影响因子：
10.5
作者：
PARKHURST, SM;ISHHOROWICZ, D
通讯作者：
ISHHOROWICZ, D
DOI：
10.1073/pnas.232688099
发表时间：
2003-01-07
影响因子：
11.1
作者：
Hall, IM;Noma, K;Grewal, SIS
通讯作者：
Grewal, SIS