Distinct subcellular localization of tRNA-derived fragments in the infective metacyclic forms of Trypanosoma cruzi

Reifur, Larissa; García-Silva, María Rosa; Poubel, Saloê Bispo; Alves, Lysangela Ronalte; Arauco, Paulo; Buiar, Diane Kelly; Goldenberg, Samuel; Cayota, Alfonso; Dallagiovanna, Bruno

doi:10.1590/s0074-02762012000600018

Cited by 24 publications

(22 citation statements)

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“…Most tRNA isoacceptors were found to be precursors of tsRNA identified in parasite-derived EVs, but specific isoacceptors were detected in higher levels as compared to others, suggesting that tsRNA are differentially packed into membrane vesicles (Table 2 and Table S3). Consistent with results published by Reifur et al [10], we found that the most abundant tsRNAs in metacyclic forms (mCell) were tRNA Glu (42%), tRNA Thr (21%), and tRNA Val (15%). Moreover, contrasting with tsRNA identified in mCell, the most abundant tsRNAs found in mVes were quite diverse and were composed of tRNA Glu (33%), tRNA Thr (17%), tRNA Gly (10%), tRNA Val (9%), and tRNA Phe (9%).…”

supporting

confidence: 92%

Characterization of the small RNA content of Trypanosoma cruzi extracellular vesicles

Bayer-Santos

Lima

Ruiz

et al. 2014

Molecular and Biochemical Parasitology

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A growing body of evidence in mammalian cells indicates that secreted vesicles can be used to mediate intercellular communication processes by transferring various bioactive molecules, including mRNAs and microRNAs. Based on these findings, we decided to analyze whether Trypanosoma cruzi-derived extracellular vesicles contain RNA molecules and performed a deep sequencing and genome-wide analysis of a size-fractioned cDNA library (16–40 nt) from extracellular vesicles secreted by noninfective epimastigote and infective metacyclic trypomastigote forms. Our data show that the small RNAs contained in these extracellular vesicles originate from multiple sources, including tRNAs. In addition, our results reveal that the variety and expression of small RNAs are different between parasite stages, suggesting diverse functions. Taken together, these observations call attention to the potential regulatory functions that these RNAs might play once transferred between parasites and/or to mammalian host cells.

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supporting

confidence: 92%

Characterization of the small RNA content of Trypanosoma cruzi extracellular vesicles

Bayer-Santos

Lima

Ruiz

et al. 2014

Molecular and Biochemical Parasitology

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“…The fraction of 5′ tRFs in both 88069 and the PiDcl1 -silenced line was somewhat higher at the three infection time points (on average 71% and 79%, respectively) than in mycelium (55% and 49%). A more dramatic change in the relative production of 5′ and 3′ tRNA halves was observed in T. cruzi , where under nutritional stress 98% of the tRFs derived from 5′ halves [ 34 ], while 87% were processed from 3′ halves in cells differentiated into the host-infective life cycle stage [ 44 ]. Differences in the proportions of 5′ mapping tRFs were observed between the isolates used in our study (90% in R0, 83% in 3928A and 55% in 88069).…”

Section: Resultsmentioning

confidence: 99%

Fragmentation of tRNA in Phytophthora infestans asexual life cycle stages and during host plant infection

et al. 2014

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BackgroundThe oomycete Phytophthora infestans possesses active RNA silencing pathways, which presumably enable this plant pathogen to control the large numbers of transposable elements present in its 240 Mb genome. Small RNAs (sRNAs), central molecules in RNA silencing, are known to also play key roles in this organism, notably in regulation of critical effector genes needed for infection of its potato host.ResultsTo identify additional classes of sRNAs in oomycetes, we mapped deep sequencing reads to transfer RNAs (tRNAs) thereby revealing the presence of 19–40 nt tRNA-derived RNA fragments (tRFs). Northern blot analysis identified abundant tRFs corresponding to half tRNA molecules. Some tRFs accumulated differentially during infection, as seen by examining sRNAs sequenced from P. infestans-potato interaction libraries. The putative connection between tRF biogenesis and the canonical RNA silencing pathways was investigated by employing hairpin RNA-mediated RNAi to silence the genes encoding P. infestans Argonaute (PiAgo) and Dicer (PiDcl) endoribonucleases. By sRNA sequencing we show that tRF accumulation is PiDcl1-independent, while Northern hybridizations detected reduced levels of specific tRNA-derived species in the PiAgo1 knockdown line.ConclusionsOur findings extend the sRNA diversity in oomycetes to include fragments derived from non-protein-coding RNA transcripts and identify tRFs with elevated levels during infection of potato by P. infestans.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-014-0308-1) contains supplementary material, which is available to authorized users.

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“…The tRNA-derived fragments of ∼32 nt in length mainly derived from the 5 arm of a restricted group of precursors tRNAs, favoring the idea of a controlled process. Shortly after, the specific production by T. cruzi of different populations of small RNAs derived not only from tRNAs and rRNAs but from sn/snoRNAs and coding genes (CDS) was also reported [12,31].…”

Section: Q2mentioning

confidence: 99%

Profiling of small RNA cargo of extracellular vesicles shed by Trypanosoma cruzi reveals a specific extracellular signature

Fernández-Calero

Garcia-Silva

Peña

et al. 2015

Molecular and Biochemical Parasitology

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Distinct subcellular localization of tRNA-derived fragments in the infective metacyclic forms of Trypanosoma cruzi

Cited by 24 publications

References 21 publications

Characterization of the small RNA content of Trypanosoma cruzi extracellular vesicles

Characterization of the small RNA content of Trypanosoma cruzi extracellular vesicles

Fragmentation of tRNA in Phytophthora infestans asexual life cycle stages and during host plant infection

Profiling of small RNA cargo of extracellular vesicles shed by Trypanosoma cruzi reveals a specific extracellular signature

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