Fragmentation of extracellular ribosomes and tRNAs shapes the extracellular RNAome.
Fragmentation of extracellular ribosomes and tRNAs shapes the extracellular RNAome.
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DOI：
10.1093/nar/gkaa674
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发表时间：
2020-12-16
影响因子：
14.9
通讯作者：
Cayota A
中科院分区：
文献类型：
--
作者：
Tosar JP;Segovia M;Castellano M;Gámbaro F;Akiyama Y;Fagúndez P;Olivera Á;Costa B;Possi T;Hill M;Ivanov P;Cayota A
大部分细胞外RNA（exRNA）不与细胞外囊泡（EV）共纯化，并且保留在细胞条件培养基或哺乳动物血清的超离心上清液中。然而，对EV以外的exRNA知之甚少。我们先前已经表明，非囊泡exRNA组分的组成高度偏向于能够形成RNA酶保护二聚体的特定tRNA衍生片段。为了解决exRNA分析中的稳定性问题，我们开发了一种基于对用RNA酶抑制剂（RI）处理的非囊泡细胞外样品的尺寸排阻色谱（SEC）级分进行测序的方法。该方法揭示了当酶促RNA降解被抑制时exRNA群体的显著组成变化。我们证明了核糖体和全长tRNA的存在下，在各种哺乳动物细胞系的细胞条件培养基。它们的片段化产生一些高度抗降解的小RNA。一些最丰富的exRNA的细胞外生物发生表明，细胞外丰度不是估计RNA分泌速率的可靠输入。最后，我们表明，含有细胞外核糖体的色谱组分可能不是沉默的免疫学的角度来看，可能被解码为损伤相关的分子模式。
A major proportion of extracellular RNAs (exRNAs) do not copurify with extracellular vesicles (EVs) and remain in ultracentrifugation supernatants of cell-conditioned medium or mammalian blood serum. However, little is known about exRNAs beyond EVs. We have previously shown that the composition of the nonvesicular exRNA fraction is highly biased toward specific tRNA-derived fragments capable of forming RNase-protecting dimers. To solve the problem of stability in exRNA analysis, we developed a method based on sequencing the size exclusion chromatography (SEC) fractions of nonvesicular extracellular samples treated with RNase inhibitors (RI). This method revealed dramatic compositional changes in exRNA population when enzymatic RNA degradation was inhibited. We demonstrated the presence of ribosomes and full-length tRNAs in cell-conditioned medium of a variety of mammalian cell lines. Their fragmentation generates some small RNAs that are highly resistant to degradation. The extracellular biogenesis of some of the most abundant exRNAs demonstrates that extracellular abundance is not a reliable input to estimate RNA secretion rates. Finally, we showed that chromatographic fractions containing extracellular ribosomes are probably not silent from an immunological perspective and could possibly be decoded as damage-associated molecular patterns.
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影响因子：
8.8
作者：
Chiou, Ni-Ting;Kageyama, Robin;Ansel, K. Mark
通讯作者：
Ansel, K. Mark
细胞外代谢能量可以促进癌症的进展。
影响因子：
64.5
作者：
Loo JM;Scherl A;Nguyen A;Man FY;Weinberg E;Zeng Z;Saltz L;Paty PB;Tavazoie SF
通讯作者：
Tavazoie SF
DOI：
10.1007/s00018-018-2842-8
发表时间：
2018-10
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Driedonks TAP;van der Grein SG;Ariyurek Y;Buermans HPJ;Jekel H;Chow FWN;Wauben MHM;Buck AH;'t Hoen PAC;Nolte-'t Hoen ENM
通讯作者：
Nolte-'t Hoen ENM
影响因子：
48
作者：
Cozen AE;Quartley E;Holmes AD;Hrabeta-Robinson E;Phizicky EM;Lowe TM
通讯作者：
Lowe TM
影响因子：
16.6
作者：
Antoury L;Hu N;Balaj L;Das S;Georghiou S;Darras B;Clark T;Breakefield XO;Wheeler TM
通讯作者：
Wheeler TM