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PMID: 20308323 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, N.I.H., Intramural Research Support, Non-U.S. Gov't

Human AlkB homolog ABH8 Is a tRNA methyltransferase required for wobble uridine modification and DNA damage survival.

Molecular and cellular biology ·Vol. 30 ·No. 10 ·2010-05-00 ·Pages 2449-59

Fu D, Brophy JA, Chan CT, Atmore KA, Begley U, Paules RS, Dedon PC, Begley TJ, Samson LD

Abstract

tRNA nucleosides are extensively modified to ensure their proper function in translation. However, many of the enzymes responsible for tRNA modifications in mammals await identification. Here, we show that human AlkB homolog 8 (ABH8) catalyzes tRNA methylation to generate 5-methylcarboxymethyl uridine (mcm(5)U) at the wobble position of certain tRNAs, a critical anticodon loop modification linked to DNA damage survival. We find that ABH8 interacts specifically with tRNAs containing mcm(5)U and that purified ABH8 complexes methylate RNA in vitro. Significantly, ABH8 depletion in human cells reduces endogenous levels of mcm(5)U in RNA and increases cellular sensitivity to DNA-damaging agents. Moreover, DNA-damaging agents induce ABH8 expression in an ATM-dependent manner. These results expand the role of mammalian AlkB proteins beyond that of direct DNA repair and support a regulatory mechanism in the DNA damage response pathway involving modulation of tRNA modification.

MeSH Terms
AlkB Homolog 8, tRNA Methyltransferase Animals Cell Line DNA Damage Humans Molecular Sequence Data Molecular Structure Nucleic Acid Conformation Phylogeny RNA, Transfer/chemistry,genetics,metabolism Uridine/chemistry,metabolism tRNA Methyltransferases/classification,genetics,metabolism
Chemicals
RNA, Transfer ALKBH8 protein, human AlkB Homolog 8, tRNA Methyltransferase tRNA Methyltransferases Uridine
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Fu Dragony
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02138, USA.
Brophy Jennifer A N
Chan Clement T Y
Atmore Kyle A
Begley Ulrike
Paules Richard S
Dedon Peter C
Begley Thomas J
Samson Leona D
References (44)
44 references, click to expand
  1. Copper-dependent cleavage of DNA by bleomycin.
    Biochemistry. 1987 Feb 10;26(3):931-42 PMID: 2436656
  2. tRNA's wobble decoding of the genome: 40 years of modification.
    J Mol Biol. 2007 Feb 9;366(1):1-13 PMID: 17187822
  3. Hot spots for modulating toxicity identified by genomic phenotyping and localization mapping.
    Mol Cell. 2004 Oct 8;16(1):117-25 PMID: 15469827
  4. A functional proteomics approach links the ubiquitin-related modifier Urm1 to a tRNA modification pathway.
    Proc Natl Acad Sci U S A. 2008 Nov 25;105(47):18255-60 PMID: 19017811
  5. HemK2 protein, encoded on human chromosome 21, methylates translation termination factor eRF1.
    FEBS Lett. 2008 Jul 9;582(16):2352-6 PMID: 18539146
  6. Sensitization of human carcinoma cells to alkylating agents by small interfering RNA suppression of 3-alkyladenine-DNA glycosylase.
    Cancer Res. 2005 Nov 15;65(22):10472-7 PMID: 16288039
  7. Cysteine dioxygenase: structure and mechanism.
    Chem Commun (Camb). 2007 Aug 28;(32):3338-49 PMID: 18019494
  8. DNA protein kinase-dependent G2 checkpoint revealed following knockdown of ataxia-telangiectasia mutated in human mammary epithelial cells.
    Cancer Res. 2008 Jan 1;68(1):89-97 PMID: 18172300
  9. The DNA-repair protein AlkB, EGL-9, and leprecan define new families of 2-oxoglutarate- and iron-dependent dioxygenases.
    Genome Biol. 2001;2(3):RESEARCH0007 PMID: 11276424
  10. The RNA methyltransferase Misu (NSun2) mediates Myc-induced proliferation and is upregulated in tumors.
    Curr Biol. 2006 May 23;16(10):971-81 PMID: 16713953
  11. Translational reprogramming following UVB irradiation is mediated by DNA-PKcs and allows selective recruitment to the polysomes of mRNAs encoding DNA repair enzymes.
    Genes Dev. 2009 May 15;23(10):1207-20 PMID: 19451221
  12. Novel methyltransferase for modified uridine residues at the wobble position of tRNA.
    Mol Cell Biol. 2003 Dec;23(24):9283-92 PMID: 14645538
  13. Ubiquitin-related modifier Urm1 acts as a sulphur carrier in thiolation of eukaryotic transfer RNA.
    Nature. 2009 Mar 12;458(7235):228-32 PMID: 19145231
  14. Phylogenomic identification of five new human homologs of the DNA repair enzyme AlkB.
    BMC Genomics. 2003 Dec 10;4(1):48 PMID: 14667252
  15. Trm9-catalyzed tRNA modifications link translation to the DNA damage response.
    Mol Cell. 2007 Dec 14;28(5):860-70 PMID: 18082610
  16. It's a mod mod tRNA world.
    Nat Chem Biol. 2008 Mar;4(3):162-4 PMID: 18277975
  17. Activities of the chaperonin containing TCP-1 (CCT): implications for cell cycle progression and cytoskeletal organisation.
    Cell Stress Chaperones. 2009 Jan;14(1):23-31 PMID: 18595008
  18. tRNA and protein methylase complexes mediate zymocin toxicity in yeast.
    Mol Microbiol. 2008 Sep;69(5):1266-77 PMID: 18657261
  19. Damage recovery pathways in Saccharomyces cerevisiae revealed by genomic phenotyping and interactome mapping.
    Mol Cancer Res. 2002 Dec;1(2):103-12 PMID: 12496357
  20. Human and bacterial oxidative demethylases repair alkylation damage in both RNA and DNA.
    Nature. 2003 Feb 20;421(6925):859-63 PMID: 12594517
  21. AlkB-mediated oxidative demethylation reverses DNA damage in Escherichia coli.
    Nature. 2002 Sep 12;419(6903):178-82 PMID: 12226668
  22. Identification of genes encoding tRNA modification enzymes by comparative genomics.
    Methods Enzymol. 2007;425:153-83 PMID: 17673083
  23. Oxidative demethylation by Escherichia coli AlkB directly reverts DNA base damage.
    Nature. 2002 Sep 12;419(6903):174-8 PMID: 12226667
  24. Probability-based validation of protein identifications using a modified SEQUEST algorithm.
    Anal Chem. 2002 Nov 1;74(21):5593-9 PMID: 12433093
  25. The obesity-associated FTO gene encodes a 2-oxoglutarate-dependent nucleic acid demethylase.
    Science. 2007 Nov 30;318(5855):1469-72 PMID: 17991826
  26. Expression and sub-cellular localization of human ABH family molecules.
    J Cell Mol Med. 2007 Sep-Oct;11(5):1105-16 PMID: 17979886
  27. Compilation of tRNA sequences and sequences of tRNA genes.
    Nucleic Acids Res. 2005 Jan 1;33(Database issue):D139-40 PMID: 15608164
  28. Repair deficient mice reveal mABH2 as the primary oxidative demethylase for repairing 1meA and 3meC lesions in DNA.
    EMBO J. 2006 May 17;25(10):2189-98 PMID: 16642038
  29. Proteome survey reveals modularity of the yeast cell machinery.
    Nature. 2006 Mar 30;440(7084):631-6 PMID: 16429126
  30. A novel human AlkB homologue, ALKBH8, contributes to human bladder cancer progression.
    Cancer Res. 2009 Apr 1;69(7):3157-64 PMID: 19293182
  31. Human telomerase and Cajal body ribonucleoproteins share a unique specificity of Sm protein association.
    Genes Dev. 2006 Mar 1;20(5):531-6 PMID: 16481465
  32. How selenium has altered our understanding of the genetic code.
    Mol Cell Biol. 2002 Jun;22(11):3565-76 PMID: 11997494
  33. Nitrogen mustard inhibits transcription and translation in a cell free system.
    Nucleic Acids Res. 1995 Sep 11;23(17):3508-15 PMID: 7567463
  34. Non-heme dioxygenases: cellular sensors and regulators jelly rolled into one?
    Nat Chem Biol. 2007 Mar;3(3):144-53 PMID: 17301803
  35. The zinc finger protein Ynr046w is plurifunctional and a component of the eRF1 methyltransferase in yeast.
    J Biol Chem. 2006 Nov 24;281(47):36140-8 PMID: 17008308
  36. Repair of alkylated DNA: recent advances.
    DNA Repair (Amst). 2007 Apr 1;6(4):429-42 PMID: 17112791
  37. Trm11p and Trm112p are both required for the formation of 2-methylguanosine at position 10 in yeast tRNA.
    Mol Cell Biol. 2005 Jun;25(11):4359-70 PMID: 15899842
  38. AlkB restores the biological function of mRNA and tRNA inactivated by chemical methylation.
    Mol Cell. 2004 Oct 8;16(1):107-16 PMID: 15469826
  39. MODOMICS: a database of RNA modification pathways. 2008 update.
    Nucleic Acids Res. 2009 Jan;37(Database issue):D118-21 PMID: 18854352
  40. AlkB reverses etheno DNA lesions caused by lipid oxidation in vitro and in vivo.
    Nat Struct Mol Biol. 2005 Oct;12(10):855-60 PMID: 16200073
  41. A new gene (alkB) of Escherichia coli that controls sensitivity to methyl methane sulfonate.
    J Bacteriol. 1983 Mar;153(3):1301-7 PMID: 6337994
  42. Methylation of tRNAAsp by the DNA methyltransferase homolog Dnmt2.
    Science. 2006 Jan 20;311(5759):395-8 PMID: 16424344
  43. Genes required for ionizing radiation resistance in yeast.
    Nat Genet. 2001 Dec;29(4):426-34 PMID: 11726929
  44. Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line.
    Nucleic Acids Res. 1990 Jun 25;18(12):3587-96 PMID: 2194165
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01604-09
Published
2010-05-00
Epub
2010-00-22
Pages
2449-59
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2863699
Subset
IM
Grants
NCI NIH HHS · R01 CA055042 · United States
NCRR NIH HHS · S10-RR023783 · United States
NCRR NIH HHS · S10 RR023783 · United States
NIEHS NIH HHS · ES01701 · United States
NCI NIH HHS · CA055042 · United States
NIEHS NIH HHS · P30 ES002109 · United States
NIEHS NIH HHS · ES002109 · United States
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1128/MCB.01604-09 PMC Full Text View journal details
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