tRNA expression and modification landscapes, and their dynamics during zebrafish embryo development

tRNA genes exist in multiple copies in the genome of all organisms across the three domains of life. Besides the sequence differences across tRNA copies, extensive post-transcriptional modification adds a further layer to tRNA diversification. Whilst the crucial role of tRNAs as adapter molecules in protein translation is well established, whether all tRNA are actually expressed, and whether the differences across isodecoders play any regulatory role is only recently being uncovered. Here we built upon recent developments in the use of NGS-based methods for RNA modification detection and developed tRAM-seq, an experimental protocol and in silico analysis pipeline to investigate tRNA expression and modification. Using tRAM-seq we analysed the full ensemble of nucleo-cytoplasmic and mitochondrial tRNAs during embryonic development of the model vertebrate zebrafish. We show that the repertoire of tRNAs changes during development, with an apparent major switch in tRNA isodecoder expression and modification profile taking place around the start of gastrulation. Taken together, our findings suggest the existence of a general reprogramming of the expressed tRNA pool, possibly gearing the translational machinery for distinct stages of the delicate and crucial process of embryo development.

[1]  Andrew Behrens,et al.  Selective gene expression maintains human tRNA anticodon pools during differentiation , 2024, Nature cell biology.

[2]  A. Pauli,et al.  eIF4E1b is a non-canonical eIF4E protecting maternal dormant mRNAs , 2023, EMBO reports.

[3]  C. Kimchi-Sarfaty,et al.  Advances in methods for tRNA sequencing and quantification. , 2023, Trends in genetics : TIG.

[4]  T. Wurdinger,et al.  ALL-tRNAseq enables robust tRNA profiling in tissue samples , 2023, Genes & development.

[5]  M. Schaefer,et al.  Identification of RNA helicases with unwinding activity on angiogenin-processed tRNAs , 2023, Nucleic acids research.

[6]  L. Qu,et al.  tModBase: deciphering the landscape of tRNA modifications and their dynamic changes from epitranscriptome data , 2022, Nucleic Acids Res..

[7]  Canquan Zhou,et al.  Developmental mRNA m5C landscape and regulatory innovations of massive m5C modification of maternal mRNAs in animals , 2022, Nature Communications.

[8]  T. Pan,et al.  tRNA modification dynamics from individual organisms to metaepitranscriptomics of microbiomes. , 2022, Molecular cell.

[9]  P. Stadler,et al.  Changes of the tRNA Modification Pattern during the Development of Dictyostelium discoideum , 2021, Non-coding RNA.

[10]  Thomas J. Begley,et al.  Quantitative mapping of the cellular small RNA landscape with AQRNA-seq , 2021, Nature Biotechnology.

[11]  Tsutomu Suzuki The expanding world of tRNA modifications and their disease relevance , 2021, Nature Reviews Molecular Cell Biology.

[12]  Andrew Behrens,et al.  High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq , 2021, Molecular cell.

[13]  M. Helm,et al.  Non-Redundant tRNA Reference Sequences for Deep Sequencing Analysis of tRNA Abundance and Epitranscriptomic RNA Modifications , 2021, Genes.

[14]  J. Coller,et al.  Quantitative tRNA-sequencing uncovers metazoan tissue-specific tRNA regulation , 2020, Nature Communications.

[15]  M. Helm,et al.  Functional characterization of the human tRNA methyltransferases TRMT10A and TRMT10B , 2020, Nucleic acids research.

[16]  Joao C. Guimaraes,et al.  A rare codon-based translational program of cell proliferation , 2020, Genome Biology.

[17]  Han Rauwerda,et al.  Maternal- and Somatic-type snoRNA Expression and Processing in Zebrafish Development , 2019, bioRxiv.

[18]  Jun Xia,et al.  RNA 5-Methylcytosine Facilitates the Maternal-to-Zygotic Transition by Preventing Maternal mRNA Decay. , 2019, Molecular cell.

[19]  L. Van Haute,et al.  NSUN2 introduces 5-methylcytosines in mammalian mitochondrial tRNAs , 2019, bioRxiv.

[20]  Tsutomu Suzuki,et al.  Mammalian NSUN2 introduces 5-methylcytidines into mitochondrial tRNAs , 2019, bioRxiv.

[21]  M. Bohnsack,et al.  Eukaryotic 5-methylcytosine (m5C) RNA Methyltransferases: Mechanisms, Cellular Functions, and Links to Disease , 2019, Genes.

[22]  A. Schleiffer,et al.  The Ly6/uPAR protein Bouncer is necessary and sufficient for species-specific fertilization , 2018, Science.

[23]  Peter F. Stadler,et al.  Accurate mapping of tRNA reads , 2018, Bioinformatics.

[24]  John S. Mattick,et al.  The RNA modification landscape in human disease , 2017, RNA.

[25]  Michael H. Schwartz,et al.  Selective Enzymatic Demethylation of N2 ,N2 -Dimethylguanosine in RNA and Its Application in High-Throughput tRNA Sequencing. , 2017, Angewandte Chemie.

[26]  L. Van Haute,et al.  Dealing with an Unconventional Genetic Code in Mitochondria: The Biogenesis and Pathogenic Defects of the 5-Formylcytosine Modification in Mitochondrial tRNAMet , 2017, Biomolecules.

[27]  Antonio J Giraldez,et al.  Codon identity regulates mRNA stability and translation efficiency during the maternal‐to‐zygotic transition , 2016, The EMBO journal.

[28]  T. Pan,et al.  tRNA base methylation identification and quantification via high-throughput sequencing , 2016, RNA.

[29]  M. Helm,et al.  Analysis of RNA modifications by liquid chromatography-tandem mass spectrometry. , 2016, Methods.

[30]  R. Stroud,et al.  Crystal Structure of the Human tRNA m(1)A58 Methyltransferase-tRNA(3)(Lys) Complex: Refolding of Substrate tRNA Allows Access to the Methylation Target. , 2015, Journal of molecular biology.

[31]  Andreas Hildebrandt,et al.  The reverse transcription signature of N-1-methyladenosine in RNA-Seq is sequence dependent , 2015, Nucleic acids research.

[32]  M. Bohnsack,et al.  NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs , 2015, RNA.

[33]  Todd M. Lowe,et al.  ARM-Seq: AlkB-facilitated RNA methylation sequencing reveals a complex landscape of modified tRNA fragments , 2015, Nature Methods.

[34]  Alan Brown,et al.  Structure of the large ribosomal subunit from human mitochondria , 2014, Science.

[35]  Sebastian M. Waszak,et al.  A Dual Program for Translation Regulation in Cellular Proliferation and Differentiation , 2014, Cell.

[36]  V. Iyer,et al.  Thermostable group II intron reverse transcriptase fusion proteins and their use in cDNA synthesis and next-generation RNA sequencing , 2013, RNA.

[37]  Alfonso Valencia,et al.  APPRIS: annotation of principal and alternative splice isoforms , 2012, Nucleic Acids Res..

[38]  Johann Holzmann,et al.  A subcomplex of human mitochondrial RNase P is a bifunctional methyltransferase—extensive moonlighting in mitochondrial tRNA biogenesis , 2012, Nucleic acids research.

[39]  Milana Frenkel-Morgenstern,et al.  Genes adopt non-optimal codon usage to generate cell cycle-dependent oscillations in protein levels , 2012, Molecular systems biology.

[40]  Steve Hoffmann,et al.  Traces of post-transcriptional RNA modifications in deep sequencing data , 2011, Biological chemistry.

[41]  Francesca Tuorto,et al.  RNA methylation by Dnmt2 protects transfer RNAs against stress-induced cleavage. , 2010, Genes & development.

[42]  A. Whitehead,et al.  Simultaneous extraction of high-quality RNA and DNA from small tissue samples. , 2009, The Journal of heredity.

[43]  Frank Lyko,et al.  RNA cytosine methylation analysis by bisulfite sequencing , 2008, Nucleic acids research.

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

[45]  Chengqi Yi,et al.  Oxidative demethylation of 3‐methylthymine and 3‐methyluracil in single‐stranded DNA and RNA by mouse and human FTO , 2008, FEBS letters.

[46]  Tao Pan,et al.  Tissue-Specific Differences in Human Transfer RNA Expression , 2006, PLoS genetics.

[47]  T. Pan,et al.  Diversity of tRNA genes in eukaryotes , 2006, Nucleic acids research.

[48]  Xiaoyu Zhang,et al.  Methylation of tRNAAsp by the DNA Methyltransferase Homolog Dnmt2 , 2006, Science.

[49]  Sergey Steinberg,et al.  Compilation of tRNA sequences and sequences of tRNA genes , 2004, Nucleic Acids Res..

[50]  Z. Zehner,et al.  Isolation and characterization of the human tRNA-(N1G37) methyltransferase (TRM5) and comparison to the Escherichia coli TrmD protein. , 2004, Biochemistry.

[51]  Magnar Bjørås,et al.  Human and bacterial oxidative demethylases repair alkylation damage in both RNA and DNA , 2003, Nature.

[52]  C. Kimmel,et al.  Stages of embryonic development of the zebrafish , 1995, Developmental dynamics : an official publication of the American Association of Anatomists.

[53]  G. M. Tener,et al.  Activity of a transfer RNA modifying enzyme during the development of Drosophila and its relationship to the su(s) locus. , 1973, Journal of molecular biology.

[54]  L. Van Haute,et al.  Detection of 5-formylcytosine in Mitochondrial Transcriptome. , 2021, Methods in molecular biology.

[55]  Sebastian A. Leidel,et al.  Analysis of codon-specific translation by ribosome profiling. , 2021, Methods in enzymology.

[56]  Christiane Branlant,et al.  Identification of modified residues in RNAs by reverse transcription-based methods. , 2007, Methods in enzymology.

[57]  P. Falnes Repair of 3-methylthymine and 1-methylguanine lesions by bacterial and human AlkB proteins. , 2004, Nucleic acids research.

[58]  L. Droogmans,et al.  Detection and quantification of modified nucleotides in RNA using thin-layer chromatography. , 2004, Methods in molecular biology.

[59]  G. Keith,et al.  2'-O-methyl-5-formylcytidine (f5Cm), a new modified nucleotide at the 'wobble' of two cytoplasmic tRNAs Leu (NAA) from bovine liver. , 1996, Nucleic acids research.