MINTmap: fast and exhaustive profiling of nuclear and mitochondrial tRNA fragments from short RNA-seq data

Transfer RNA fragments (tRFs) are an established class of constitutive regulatory molecules that arise from precursor and mature tRNAs. RNA deep sequencing (RNA-seq) has greatly facilitated the study of tRFs. However, the repeat nature of the tRNA templates and the idiosyncrasies of tRNA sequences necessitate the development and use of methodologies that differ markedly from those used to analyze RNA-seq data when studying microRNAs (miRNAs) or messenger RNAs (mRNAs). Here we present MINTmap (for MItochondrial and Nuclear TRF mapping), a method and a software package that was developed specifically for the quick, deterministic and exhaustive identification of tRFs in short RNA-seq datasets. In addition to identifying them, MINTmap is able to unambiguously calculate and report both raw and normalized abundances for the discovered tRFs. Furthermore, to ensure specificity, MINTmap identifies the subset of discovered tRFs that could be originating outside of tRNA space and flags them as candidate false positives. Our comparative analysis shows that MINTmap exhibits superior sensitivity and specificity to other available methods while also being exceptionally fast. The MINTmap codes are available through https://github.com/TJU-CMC-Org/MINTmap/ under an open source GNU GPL v3.0 license.

[1]  Megumi Shigematsu,et al.  Transfer RNA as a source of small functional RNA. , 2014, Journal of molecular biology and molecular imaging.

[2]  Gyorgy Hutvagner,et al.  tRNA-Derived Fragments (tRFs): Emerging New Roles for an Ancient RNA in the Regulation of Gene Expression , 2015, Life.

[3]  N. Polacek,et al.  tRNA-Derived Fragments Target the Ribosome and Function as Regulatory Non-Coding RNA in Haloferax volcanii , 2012, Archaea.

[4]  Lisa Fish,et al.  Endogenous tRNA-Derived Fragments Suppress Breast Cancer Progression via YBX1 Displacement , 2015, Cell.

[5]  Andrey Grigoriev,et al.  Age-driven modulation of tRNA-derived fragments in Drosophila and their potential targets , 2015, Biology Direct.

[6]  H. Inokuchi,et al.  tRNADB-CE: tRNA gene database well-timed in the era of big sequence data , 2014, Front. Genet..

[7]  Yuan Chang,et al.  Extensive terminal and asymmetric processing of small RNAs from rRNAs, snoRNAs, snRNAs, and tRNAs , 2012, Nucleic acids research.

[8]  Xavier Estivill,et al.  Evidence for the biogenesis of more than 1,000 novel human microRNAs , 2014, Genome Biology.

[9]  Jie Wu,et al.  tRF2Cancer: A web server to detect tRNA-derived small RNA fragments (tRFs) and their expression in multiple cancers , 2016, Nucleic Acids Res..

[10]  A. Hopper,et al.  tRNA biology charges to the front. , 2010, Genes & development.

[11]  Patricia P. Chan,et al.  GtRNAdb 2.0: an expanded database of transfer RNA genes identified in complete and draft genomes , 2015, Nucleic Acids Res..

[12]  J. Jackman,et al.  Life without post-transcriptional addition of G−1: two alternatives for tRNAHis identity in Eukarya , 2015, RNA.

[13]  S. Yamashita,et al.  Molecular mechanisms of template-independent RNA polymerization by tRNA nucleotidyltransferases , 2014, Front. Genet..

[14]  David I. K. Martin,et al.  Deep Sequencing of Serum Small RNAs Identifies Patterns of 5′ tRNA Half and YRNA Fragment Expression Associated with Breast Cancer , 2014, Biomarkers in cancer.

[15]  S. S. Ajay,et al.  Identification and functional characterization of tRNA-derived RNA fragments (tRFs) in respiratory syncytial virus infection. , 2013, Molecular therapy : the journal of the American Society of Gene Therapy.

[16]  Xiaoyong Bao,et al.  Respiratory Syncytial Virus Utilizes a tRNA Fragment to Suppress Antiviral Responses Through a Novel Targeting Mechanism. , 2015, Molecular therapy : the journal of the American Society of Gene Therapy.

[17]  Aristeidis G Telonis,et al.  Mitochondrial tRNA-lookalikes in nuclear chromosomes: Could they be functional? , 2015, RNA biology.

[18]  Youri Hoogstrate,et al.  A comprehensive repertoire of tRNA-derived fragments in prostate cancer , 2016, Oncotarget.

[19]  Peter F. Stadler,et al.  tRNAdb 2009: compilation of tRNA sequences and tRNA genes , 2008, Nucleic Acids Res..

[20]  S. Grewal Why should cancer biologists care about tRNAs? tRNA synthesis, mRNA translation and the control of growth. , 2015, Biochimica et biophysica acta.

[21]  Oliver J. Rando,et al.  Biogenesis and function of tRNA fragments during sperm maturation and fertilization in mammals , 2016, Science.

[22]  Ilka U. Heinemann,et al.  tRNAHis-guanylyltransferase establishes tRNAHis identity , 2011, Nucleic acids research.

[23]  A. Malhotra,et al.  A novel class of small RNAs: tRNA-derived RNA fragments (tRFs). , 2009, Genes & development.

[24]  S. Goldenberg,et al.  Distinct subcellular localization of tRNA-derived fragments in the infective metacyclic forms of Trypanosoma cruzi. , 2012, Memorias do Instituto Oswaldo Cruz.

[25]  Markus Glatzel,et al.  CLP1 links tRNA metabolism to progressive motor-neuron loss , 2013, Nature.

[26]  Pedro G. Ferreira,et al.  Transcriptome and genome sequencing uncovers functional variation in humans , 2013, Nature.

[27]  Annette S. Kim,et al.  Transfer RNA detection by small RNA deep sequencing and disease association with myelodysplastic syndromes , 2015, BMC Genomics.

[28]  T. Speed,et al.  A Tetrahymena Piwi bound to mature tRNA 3' fragments activates the exonuclease Xrn2 for RNA processing in the nucleus. , 2012, Molecular cell.

[29]  Pankaj Kumar,et al.  tRFdb: a database for transfer RNA fragments , 2014, Nucleic Acids Res..

[30]  Yong Sun Lee,et al.  Compartmentalized, functional role of angiogenin during spotted fever group rickettsia-induced endothelial barrier dysfunction: evidence of possible mediation by host tRNA-derived small noncoding RNAs , 2013, BMC Infectious Diseases.

[31]  Megumi Shigematsu,et al.  tRNA-Derived Short Non-coding RNA as Interacting Partners of Argonaute Proteins , 2015, Gene regulation and systems biology.

[32]  Yi Jing,et al.  Dissecting tRNA-derived fragment complexities using personalized transcriptomes reveals novel fragment classes and unexpected dependencies , 2015, Oncotarget.

[33]  M. Ibba,et al.  tRNAs as regulators of biological processes , 2014, Front. Genet..

[34]  Robert Blelloch,et al.  Mouse ES cells express endogenous shRNAs, siRNAs, and other Microprocessor-independent, Dicer-dependent small RNAs. , 2008, Genes & development.

[35]  Praveen Sethupathy,et al.  tDRmapper: challenges and solutions to mapping, naming, and quantifying tRNA-derived RNAs from human small RNA-sequencing data , 2015, BMC Bioinformatics.

[36]  Pavel Ivanov,et al.  Angiogenin-induced tRNA-derived Stress-induced RNAs Promote Stress-induced Stress Granule Assembly* , 2010, The Journal of Biological Chemistry.

[37]  Phillipe Loher,et al.  Nuclear and mitochondrial tRNA-lookalikes in the human genome , 2014, Front. Genet..

[38]  S. Le,et al.  Pyrosequencing of small non-coding RNAs in HIV-1 infected cells: evidence for the processing of a viral-cellular double-stranded RNA hybrid , 2009, Nucleic acids research.

[39]  Masaru Tomita,et al.  Precise mapping and dynamics of tRNA-derived fragments (tRFs) in the development of Triops cancriformis (tadpole shrimp) , 2015, BMC Genetics.

[40]  Computational analysis, biochemical purification, and detection of tRNA-derived small RNA fragments. , 2014, Methods in molecular biology.

[41]  Isidore Rigoutsos,et al.  MINTbase: a framework for the interactive exploration of mitochondrial and nuclear tRNA fragments , 2016, Bioinform..

[42]  Isidore Rigoutsos,et al.  Consequential considerations when mapping tRNA fragments , 2016, BMC Bioinformatics.

[43]  D. Haussecker,et al.  Human tRNA-derived small RNAs in the global regulation of RNA silencing. , 2010, RNA.

[44]  Ya-Ming Hou,et al.  CCA addition to tRNA: Implications for tRNA quality control , 2010, IUBMB life.

[45]  S. Ferrari,et al.  Author contributions , 2021 .

[46]  Steven P Gygi,et al.  Angiogenin-induced tRNA fragments inhibit translation initiation. , 2011, Molecular cell.

[47]  Phillipe Loher,et al.  Sex hormone-dependent tRNA halves enhance cell proliferation in breast and prostate cancers , 2015, Proceedings of the National Academy of Sciences.

[48]  Patricia P. Chan,et al.  GtRNAdb: a database of transfer RNA genes detected in genomic sequence , 2008, Nucleic Acids Res..

[49]  Jordan Anaya,et al.  Meta-analysis of tRNA derived RNA fragments reveals that they are evolutionarily conserved and associate with AGO proteins to recognize specific RNA targets , 2014, BMC Biology.

[50]  R. Sachidanandam,et al.  A growth-essential Tetrahymena Piwi protein carries tRNA fragment cargo. , 2010, Genes & development.

[51]  Yi Jing,et al.  Beyond the one-locus-one-miRNA paradigm: microRNA isoforms enable deeper insights into breast cancer heterogeneity , 2015, Nucleic acids research.

[52]  Joern Pütz,et al.  Mamit-tRNA, a database of mammalian mitochondrial tRNA primary and secondary structures. , 2007, RNA.

[53]  Fedor V. Karginov,et al.  Developmentally regulated cleavage of tRNAs in the bacterium Streptomyces coelicolor , 2007, Nucleic acids research.

[54]  G. Barton,et al.  Filtering of deep sequencing data reveals the existence of abundant Dicer-dependent small RNAs derived from tRNAs. , 2009, RNA.

[55]  Lucian Ilie,et al.  SHRiMP2: Sensitive yet Practical Short Read Mapping , 2011, Bioinform..

[56]  Andrea Califano,et al.  tRNA-derived microRNA modulates proliferation and the DNA damage response and is down-regulated in B cell lymphoma , 2013, Proceedings of the National Academy of Sciences.

[57]  Praveen Sethupathy,et al.  Small tRNA-derived RNAs are increased and more abundant than microRNAs in chronic hepatitis B and C , 2015, Scientific Reports.

[58]  Yan Li,et al.  Both endo-siRNAs and tRNA-derived small RNAs are involved in the differentiation of primitive eukaryote Giardia lamblia , 2014, Proceedings of the National Academy of Sciences.

[59]  Noah Spies,et al.  Tramp-mediated Rna Surveillance Prevents Spurious Entry of Rnas into the Schizosaccharomyces Pombe Sirna Pathway Nih Public Access Author Manuscript Gene-specific Srnas Methods Fission Yeast Strains and Plasmids Generation of Small Rna Libraries for 454 Deep Sequencing Supplementary Material Acknowl , 2022 .

[60]  M. Mörl,et al.  tRNA nucleotidyltransferases: ancient catalysts with an unusual mechanism of polymerization , 2010, Cellular and Molecular Life Sciences.

[61]  Phillipe Loher,et al.  IsomiR expression profiles in human lymphoblastoid cell lines exhibit population and gender dependencies , 2014, Oncotarget.

[62]  Richard Durbin,et al.  Sequence analysis Fast and accurate short read alignment with Burrows – Wheeler transform , 2009 .

[63]  S. Yamasaki,et al.  Angiogenin cleaves tRNA and promotes stress-induced translational repression , 2009, The Journal of cell biology.

[64]  M. Hudson Human , 2018, Critical Theory and the Classical World.

[65]  Ana Kozomara,et al.  miRBase: integrating microRNA annotation and deep-sequencing data , 2010, Nucleic Acids Res..

[66]  Yi Jing,et al.  Analysis of 13 cell types reveals evidence for the expression of numerous novel primate- and tissue-specific microRNAs , 2015, Proceedings of the National Academy of Sciences.

[67]  C. Mayr,et al.  Widespread Shortening of 3′UTRs by Alternative Cleavage and Polyadenylation Activates Oncogenes in Cancer Cells , 2009, Cell.

[68]  N. Polacek,et al.  Slicing tRNAs to boost functional ncRNA diversity , 2013, RNA biology.

[69]  Brian A. Smith,et al.  tRNAHis guanylyltransferase (THG1), a unique 3′-5′ nucleotidyl transferase, shares unexpected structural homology with canonical 5′-3′ DNA polymerases , 2010, Proceedings of the National Academy of Sciences.