Genome-wide identification and characterization of tRNA-derived RNA fragments in land plants

Key messageThe manuscript by Alves et al. entitled “Genome-wide identification and characterization of tRNA-derived RNA fragments in land plants” describes the identification and characterization of tRNAderived sRNA fragments in plants. By combining bioinformatic analysis and genetic and molecular approaches, we show that tRF biogenesis does not rely on canonical microRNA/siRNA processing machinery (i.e., independent of DICER-LIKE proteins). Moreover, we provide evidences that the Arabidopsis S-like Ribonuclease 1 (RNS1) might be involved in the biogenesis of tRFs. Detailed analyses showed that plant tRFs are sorted into different types of ARGONAUTE proteins and that they have potential target candidate genes. Our work advances the understanding of the tRF biology in plants by providing evidences that plant and animal tRFs shared common features and raising the hypothesis that an interplay between tRFs and other sRNAs might be important to fine-tune gene expression and protein biosynthesis in plant cells.AbstractSmall RNA (sRNA) fragments derived from tRNAs (3′-loop, 5′-loop, anti-codon loop), named tRFs, have been reported in several organisms, including humans and plants. Although they may interfere with gene expression, their biogenesis and biological functions in plants remain poorly understood. Here, we capitalized on small RNA sequencing data from distinct species such as Arabidopsis thaliana, Oryza sativa, and Physcomitrella patens to examine the diversity of plant tRFs and provide insight into their properties. In silico analyzes of 19 to 25-nt tRFs derived from 5′ (tRF-5s) and 3′CCA (tRF-3s) tRNA loops in these three evolutionary distant species showed that they are conserved and their abundance did not correlate with the number of genomic copies of the parental tRNAs. Moreover, tRF-5 is the most abundant variant in all three species. In silico and in vivo expression analyses unraveled differential accumulation of tRFs in Arabidopsis tissues/organs, suggesting that they are not byproducts of tRNA degradation. We also verified that the biogenesis of most Arabidopsis 19–25 nt tRF-5s and tRF-3s is not primarily dependent on DICER-LIKE proteins, though they seem to be associated with ARGONAUTE proteins and have few potential targets. Finally, we provide evidence that Arabidopsis ribonuclease RNS1 might be involved in the processing and/or degradation of tRFs. Our data support the notion that an interplay between tRFs and other sRNAs might be important to fine tune gene expression and protein biosynthesis in plant cells.

[1]  P. Green,et al.  The Arabidopsis ribonuclease gene RNS1 is tightly controlled in response to phosphate limitation. , 1994, The Plant journal : for cell and molecular biology.

[2]  Xin Niu,et al.  tRNA-Derived Small Non-Coding RNAs in Response to Ischemia Inhibit Angiogenesis , 2016, Scientific Reports.

[3]  Po-Jung Huang,et al.  A Comprehensive Expression Profile of MicroRNAs and Other Classes of Non-Coding Small RNAs in Barley Under Phosphorous-Deficient and -Sufficient Conditions , 2012, DNA research : an international journal for rapid publication of reports on genes and genomes.

[4]  N. Fedoroff,et al.  The RNA-binding proteins HYL1 and SE promote accurate in vitro processing of pri-miRNA by DCL1 , 2008, Proceedings of the National Academy of Sciences.

[5]  Rogerio Margis,et al.  Description of plant tRNA-derived RNA fragments (tRFs) associated with argonaute and identification of their putative targets , 2013, Biology Direct.

[6]  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.

[7]  D. Chitwood,et al.  Organ polarity in plants is specified through the opposing activity of two distinct small regulatory RNAs. , 2006, Cold Spring Harbor symposia on quantitative biology.

[8]  E. F. Walton,et al.  Plant Methods Protocol: a Highly Sensitive Rt-pcr Method for Detection and Quantification of Micrornas , 2022 .

[9]  S. Whisson,et al.  Fragmentation of tRNA in Phytophthora infestans asexual life cycle stages and during host plant infection , 2014, BMC Microbiology.

[10]  H. Vaucheret,et al.  The Nuclear dsRNA Binding Protein HYL1 Is Required for MicroRNA Accumulation and Plant Development, but Not Posttranscriptional Transgene Silencing , 2004, Current Biology.

[11]  M. Bevan,et al.  Sugar and ABA response pathways and the control of gene expression. , 2006, Plant, cell & environment.

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

[13]  Gregory J. Hannon,et al.  Sorting of Small RNAs into Arabidopsis Argonaute Complexes Is Directed by the 5′ Terminal Nucleotide , 2008, Cell.

[14]  P. Green,et al.  Impact of transcriptional, ABA-dependent, and ABA-independent pathways on wounding regulation of RNS1 expression , 2008, Molecular Genetics and Genomics.

[15]  Manuel A. S. Santos,et al.  Conserved and highly expressed tRNA derived fragments in zebrafish , 2015, BMC Molecular Biology.

[16]  W. Du,et al.  The disturbance of small RNA pathways enhanced abscisic acid response and multiple stress responses in Arabidopsis. , 2008, Plant, cell & environment.

[17]  Yong Sun Lee,et al.  Small Non-coding Transfer RNA-Derived RNA Fragments (tRFs): Their Biogenesis, Function and Implication in Human Diseases , 2015, Genomics & informatics.

[18]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[19]  Ilha Lee,et al.  SUPPRESSOR OF FRIGIDA4, Encoding a C2H2-Type Zinc Finger Protein, Represses Flowering by Transcriptional Activation of Arabidopsis FLOWERING LOCUS C[W] , 2006, The Plant Cell Online.

[20]  Edwards Allen,et al.  DICER-LIKE 4 functions in trans-acting small interfering RNA biogenesis and vegetative phase change in Arabidopsis thaliana. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Thomas Tuschl,et al.  The growing catalog of small RNAs and their association with distinct Argonaute/Piwi family members , 2008, Development.

[22]  S. Luo,et al.  Global identification of microRNA–target RNA pairs by parallel analysis of RNA ends , 2008, Nature Biotechnology.

[23]  A. Hopper,et al.  A decade of surprises for tRNA nuclear-cytoplasmic dynamics. , 2008, Trends in cell biology.

[24]  T. Steitz,et al.  A story with a good ending: tRNA 3'-end maturation by CCA-adding enzymes. , 2006, Current opinion in structural biology.

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

[26]  J. Yong,et al.  tRNA binds to cytochrome c and inhibits caspase activation. , 2010, Molecular cell.

[27]  Gene W. Yeo,et al.  Blurred Boundaries: The RNA Binding Protein Lin28A Is Also an Epigenetic Regulator. , 2016, Molecular cell.

[28]  Paulo Mazzafera,et al.  An Arabidopsis Mitochondrial Uncoupling Protein Confers Tolerance to Drought and Salt Stress in Transgenic Tobacco Plants , 2011, PloS one.

[29]  N. Fedoroff,et al.  Dynamic Regulation of ARGONAUTE4 within Multiple Nuclear Bodies in Arabidopsis thaliana , 2008, PLoS genetics.

[30]  C. Matiolli,et al.  Involvement of microRNA-related regulatory pathways in the glucose-mediated control of Arabidopsis early seedling development , 2013, Journal of experimental botany.

[31]  G. Hutvagner,et al.  Small RNAs derived from the 5′ end of tRNA can inhibit protein translation in human cells , 2013, RNA biology.

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

[33]  S. Asha,et al.  Transfer RNA Derived Small RNAs Targeting Defense Responsive Genes Are Induced during Phytophthora capsici Infection in Black Pepper (Piper nigrum L.) , 2016, Front. Plant Sci..

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

[35]  Filip Rolland,et al.  Role of the Arabidopsis Glucose Sensor HXK1 in Nutrient, Light, and Hormonal Signaling , 2003, Science.

[36]  Sang Yeol Lee,et al.  MED18 interaction with distinct transcription factors regulates multiple plant functions , 2014, Nature Communications.

[37]  Wen-Hsiung Li,et al.  Uncovering Small RNA-Mediated Responses to Phosphate Deficiency in Arabidopsis by Deep Sequencing1[W][OA] , 2009, Plant Physiology.

[38]  John P Sumpter,et al.  Populations of a cyprinid fish are self-sustaining despite widespread feminization of males , 2014, BMC Biology.

[39]  A. Telonis,et al.  Four-leaf clover qRT-PCR: A convenient method for selective quantification of mature tRNA , 2015, RNA biology.

[40]  Y. Qi,et al.  A Dicer-Independent Route for Biogenesis of siRNAs that Direct DNA Methylation in Arabidopsis. , 2016, Molecular cell.

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

[42]  Peter R. Crane,et al.  The origin and early evolution of plants on land , 1997, Nature.

[43]  Y. Guisez,et al.  Oxidative stress-related responses at transcriptional and enzymatic levels after exposure to Cd or Cu in a multipollution context. , 2009, Journal of plant physiology.

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

[45]  Sean R. Davis,et al.  NCBI GEO: archive for functional genomics data sets—update , 2012, Nucleic Acids Res..

[46]  Lex E. Flagel,et al.  RNase T2 genes from rice and the evolution of secretory ribonucleases in plants , 2010, Molecular Genetics and Genomics.

[47]  R. L. Hurto Unexpected functions of tRNA and tRNA processing enzymes. , 2011, Advances in experimental medicine and biology.

[48]  I. Henderson,et al.  Dissecting Arabidopsis thaliana DICER function in small RNA processing, gene silencing and DNA methylation patterning , 2006, Nature Genetics.

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

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

[51]  M. Nei,et al.  MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. , 2011, Molecular biology and evolution.

[52]  Daniel Gautheret,et al.  Genome-wide discovery and analysis of microRNAs and other small RNAs from rice embryogenic callus , 2011, RNA biology.

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

[54]  Patrick Xuechun Zhao,et al.  psRNATarget: a plant small RNA target analysis server , 2011, Nucleic Acids Res..

[55]  David Tollervey,et al.  RNA in pieces. , 2011, Trends in genetics : TIG.

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

[57]  R. Klose,et al.  Understanding the relationship between DNA methylation and histone lysine methylation , 2014, Biochimica et biophysica acta.

[58]  Kristof Engelen,et al.  Insights into the Role of the Berry-Specific Ethylene Responsive Factor VviERF045 , 2016, Frontiers in plant science.

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

[60]  M. Helm,et al.  Detection of RNA modifications , 2010, RNA biology.

[61]  R. Sachidanandam,et al.  Bacterial argonaute samples the transcriptome to identify foreign DNA. , 2013, Molecular cell.

[62]  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 .

[63]  T. Schedl,et al.  RNA-binding proteins. , 2006, WormBook : the online review of C. elegans biology.

[64]  Pamela J Green,et al.  tRNA cleavage is a conserved response to oxidative stress in eukaryotes. , 2008, RNA.

[65]  T. Pan,et al.  Diversity of human tRNA genes from the 1000-genomes project , 2013, RNA biology.

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

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

[68]  F. Nogueira,et al.  Global analysis of the sugarcane microtranscriptome reveals a unique composition of small RNAs associated with axillary bud outgrowth , 2013, Journal of experimental botany.

[69]  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.

[70]  M. Prigge,et al.  Evolutionary crossroads in developmental biology: Physcomitrella patens , 2010, Development.

[71]  Pengfei Cai,et al.  A Deep Analysis of the Small Non-Coding RNA Population in Schistosoma japonicum Eggs , 2013, PloS one.

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

[73]  Adam M. Gustafson,et al.  Genetic and Functional Diversification of Small RNA Pathways in Plants , 2004, PLoS biology.