Tags

Type your tag names separated by a space and hit enter

Dimerization confers increased stability to nucleases in 5' halves from glycine and glutamic acid tRNAs.
Nucleic Acids Res. 2018 09 28; 46(17):9081-9093.NA

Abstract

We have previously shown that 5' halves from tRNAGlyGCC and tRNAGluCUC are the most enriched small RNAs in the extracellular space of human cell lines, and especially in the non-vesicular fraction. Extracellular RNAs are believed to require protection by either encapsulation in vesicles or ribonucleoprotein complex formation. However, deproteinization of non-vesicular tRNA halves does not affect their retention in size-exclusion chromatography. Thus, we considered alternative explanations for their extracellular stability. In-silico analysis of the sequence of these tRNA-derived fragments showed that tRNAGly 5' halves can form homodimers or heterodimers with tRNAGlu 5' halves. This capacity is virtually unique to glycine tRNAs. By analyzing synthetic oligonucleotides by size exclusion chromatography, we provide evidence that dimerization is possible in vitro. tRNA halves with single point substitutions preventing dimerization are degraded faster both in controlled nuclease digestion assays and after transfection in cells, showing that dimerization can stabilize tRNA halves against the action of cellular nucleases. Finally, we give evidence supporting dimerization of endogenous tRNAGlyGCC 5' halves inside cells. Considering recent reports have shown that 5' tRNA halves from Ala and Cys can form tetramers, our results highlight RNA intermolecular structures as a new layer of complexity in the biology of tRNA-derived fragments.

Authors+Show Affiliations

Functional Genomics Unit, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay. Nuclear Research Center, Faculty of Science, Universidad de la República, Montevideo 11400, Uruguay.Functional Genomics Unit, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay. Nuclear Research Center, Faculty of Science, Universidad de la República, Montevideo 11400, Uruguay.Functional Genomics Unit, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay. Group of Biomolecular Simulations, Institut Pasteur de Montevideo. Montevideo 11400, Uruguay.Group of Biomolecular Simulations, Institut Pasteur de Montevideo. Montevideo 11400, Uruguay.Architecture et Réactivité de l'ARN, Université de Strasbourg, Institut de biologie moléculaire et cellulaire du CNRS, 15 rue René Descartes, 67084 Strasbourg, France.Functional Genomics Unit, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay. Department of Medicine, Faculty of Medicine, Universidad de la República, Montevideo 11600, Uruguay.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29893896

Citation

Tosar, Juan Pablo, et al. "Dimerization Confers Increased Stability to Nucleases in 5' Halves From Glycine and Glutamic Acid TRNAs." Nucleic Acids Research, vol. 46, no. 17, 2018, pp. 9081-9093.
Tosar JP, Gámbaro F, Darré L, et al. Dimerization confers increased stability to nucleases in 5' halves from glycine and glutamic acid tRNAs. Nucleic Acids Res. 2018;46(17):9081-9093.
Tosar, J. P., Gámbaro, F., Darré, L., Pantano, S., Westhof, E., & Cayota, A. (2018). Dimerization confers increased stability to nucleases in 5' halves from glycine and glutamic acid tRNAs. Nucleic Acids Research, 46(17), 9081-9093. https://doi.org/10.1093/nar/gky495
Tosar JP, et al. Dimerization Confers Increased Stability to Nucleases in 5' Halves From Glycine and Glutamic Acid TRNAs. Nucleic Acids Res. 2018 09 28;46(17):9081-9093. PubMed PMID: 29893896.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Dimerization confers increased stability to nucleases in 5' halves from glycine and glutamic acid tRNAs. AU - Tosar,Juan Pablo, AU - Gámbaro,Fabiana, AU - Darré,Leonardo, AU - Pantano,Sergio, AU - Westhof,Eric, AU - Cayota,Alfonso, PY - 2018/02/06/received PY - 2018/05/22/accepted PY - 2018/6/13/pubmed PY - 2019/8/14/medline PY - 2018/6/13/entrez SP - 9081 EP - 9093 JF - Nucleic acids research JO - Nucleic Acids Res. VL - 46 IS - 17 N2 - We have previously shown that 5' halves from tRNAGlyGCC and tRNAGluCUC are the most enriched small RNAs in the extracellular space of human cell lines, and especially in the non-vesicular fraction. Extracellular RNAs are believed to require protection by either encapsulation in vesicles or ribonucleoprotein complex formation. However, deproteinization of non-vesicular tRNA halves does not affect their retention in size-exclusion chromatography. Thus, we considered alternative explanations for their extracellular stability. In-silico analysis of the sequence of these tRNA-derived fragments showed that tRNAGly 5' halves can form homodimers or heterodimers with tRNAGlu 5' halves. This capacity is virtually unique to glycine tRNAs. By analyzing synthetic oligonucleotides by size exclusion chromatography, we provide evidence that dimerization is possible in vitro. tRNA halves with single point substitutions preventing dimerization are degraded faster both in controlled nuclease digestion assays and after transfection in cells, showing that dimerization can stabilize tRNA halves against the action of cellular nucleases. Finally, we give evidence supporting dimerization of endogenous tRNAGlyGCC 5' halves inside cells. Considering recent reports have shown that 5' tRNA halves from Ala and Cys can form tetramers, our results highlight RNA intermolecular structures as a new layer of complexity in the biology of tRNA-derived fragments. SN - 1362-4962 UR - https://www.unboundmedicine.com/medline/citation/29893896/Dimerization_confers_increased_stability_to_nucleases_in_5'_halves_from_glycine_and_glutamic_acid_tRNAs_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gky495 DB - PRIME DP - Unbound Medicine ER -