Arabidopsis apoplastic fluid contains sRNA- and circular RNA-protein complexes that are located outside extracellular vesicles.
Arabidopsis apoplastic fluid contains sRNA- and circular RNA-protein complexes that are located outside extracellular vesicles.
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DOI：
10.1093/plcell/koac043
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发表时间：
2022-04-26
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影响因子：
--
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中科院分区：
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此前，我们已经证明从拟南芥叶中纯化的质外体洗涤液（AWF）含有小RNA（sRNA）。为了研究这些sRNA是否被封装在细胞外囊泡（EV）内，我们用蛋白酶胰蛋白酶和RNase A处理从拟南芥叶中分离的EV，其应该降解位于EV外部的RNA，而不是位于EV内部的RNA。这些分析表明，质外体RNA主要位于外部，并与蛋白质相关。对这些细胞外RNA（exRNA）的进一步分析显示，它们包括sRNA和长非编码RNA（lncRNA），包括环状RNA（circRNA）。我们还发现exRNA在转录后修饰N6-甲基腺嘌呤（m6 A）中高度富集。与此一致，我们在AWF中鉴定了一种推定的m6 A结合蛋白，即富含甘氨酸的RNA结合蛋白7（GRP 7），以及sRNA结合蛋白Argonaute 2（AGO 2）。这两种蛋白质与lncRNA（包括circRNA）共免疫沉淀。GRP 7或AGO 2的突变引起AWF的sRNA和lncRNA含量的变化，表明这些蛋白质有助于exRNA的分泌和/或稳定。我们认为，位于电动汽车外部的exRNA介导了宿主诱导的基因沉默，而不是位于电动汽车内部的RNA。拟南芥叶片的质外体含有多种小而长的非编码RNA，包括环状RNA，它们与蛋白质复合物结合并位于细胞外囊泡之外。
Previously, we have shown that apoplastic wash fluid (AWF) purified from Arabidopsis leaves contains small RNAs (sRNAs). To investigate whether these sRNAs are encapsulated inside extracellular vesicles (EVs), we treated EVs isolated from Arabidopsis leaves with the protease trypsin and RNase A, which should degrade RNAs located outside EVs but not those located inside. These analyses revealed that apoplastic RNAs are mostly located outside and are associated with proteins. Further analyses of these extracellular RNAs (exRNAs) revealed that they include both sRNAs and long noncoding RNAs (lncRNAs), including circular RNAs (circRNAs). We also found that exRNAs are highly enriched in the posttranscriptional modification N6-methyladenine (m6A). Consistent with this, we identified a putative m6A-binding protein in AWF, GLYCINE-RICH RNA-BINDING PROTEIN 7 (GRP7), as well as the sRNA-binding protein ARGONAUTE2 (AGO2). These two proteins coimmunoprecipitated with lncRNAs, including circRNAs. Mutation of GRP7 or AGO2 caused changes in both the sRNA and lncRNA content of AWF, suggesting that these proteins contribute to the secretion and/or stabilization of exRNAs. We propose that exRNAs located outside of EVs mediate host-induced gene silencing, rather than RNA located inside EVs. The apoplast of Arabidopsis leaves contains diverse small and long-noncoding RNAs, including circular RNAs, that are bound to protein complexes and are located outside extracellular vesicles.
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影响因子：
3.7
作者：
Harvey JJ;Lewsey MG;Patel K;Westwood J;Heimstädt S;Carr JP;Baulcombe DC
通讯作者：
Baulcombe DC
DOI：
10.1007/978-981-13-1426-1_7
发表时间：
2018-01-01
期刊：
CIRCULAR RNAS: BIOGENESIS AND FUNCTIONS
影响因子：
--
作者：
Bose, Rumela;Ain, Rupasri
通讯作者：
Ain, Rupasri
影响因子：
64.8
作者：
Fu, Zheng Qing;Guo, Ming;Alfano, James R.
通讯作者：
Alfano, James R.
DOI：
10.1126/science.aar4142
发表时间：
2018-06-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Cai Q;Qiao L;Wang M;He B;Lin FM;Palmquist J;Huang SD;Jin H
通讯作者：
Jin H
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y