title: Identification of a novel methyltransferase, Bmt2, responsible for the N-1-methyl-adenosine base modification of 25S rRNA in Saccharomyces cerevisiae.
abstract: The 25S rRNA of yeast contains several base modifications in the functionally important regions. The enzymes responsible for most of these base modifications remained unknown. Recently, we identified Rrp8 as a methyltransferase involved in m(1)A645 modification of 25S rRNA. Here, we discovered a previously uncharacterized gene YBR141C to be responsible for second m(1)A2142 modification of helix 65 of 25S rRNA. The gene was identified by reversed phase-HPLC screening of all deletion mutants of putative RNA methyltransferase and was confirmed by gene complementation and phenotypic characterization. Because of the function of its encoded protein, YBR141C was named BMT2 (base methyltransferase of 25S RNA). Helix 65 belongs to domain IV, which accounts for most of the intersubunit surface of the large subunit. The 3D structure prediction of Bmt2 supported it to be an Ado Met methyltransferase belonging to Rossmann fold superfamily. In addition, we demonstrated that the substitution of G180R in the S-adenosyl-L-methionine-binding motif drastically reduces the catalytic function of the protein in vivo. Furthermore, we analysed the significance of m(1)A2142 modification in ribosome synthesis and translation. Intriguingly, the loss of m(1)A2142 modification confers anisomycin and peroxide sensitivity to the cells. Our results underline the importance of RNA modifications in cellular physiology.
authors.affiliation: Institute of Molecular Biosciences, Goethe University Frankfurt 60438, Max-von-Laue Street 9, 60438 Frankfurt/M, Germany.
authors.forename: Sunny
authors.initials: S
authors.lastname: Sharma
authors.forename: Peter
authors.initials: P
authors.lastname: Watzinger
authors.forename: Peter
authors.initials: P
authors.lastname: Kötter
authors.forename: Karl-Dieter
authors.initials: KD
authors.lastname: Entian
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000633:toxicity; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D014176:Protein Biosynthesis; D012335:RNA, Ribosomal / Q000737:chemistry / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D012335:RNA, Ribosomal; D029701:Saccharomyces cerevisiae Proteins; C065039:RNA, ribosomal, 25S; D006861:Hydrogen Peroxide; C582433:Bmt2 protein, S cerevisiae; D008780:Methyltransferases; D000241:Adenosine
references.citation: Nucleic Acids Res. 2008 Jul 1;36(Web Server issue):W197-201
references.pmid: 18463136
references.citation: Nucleic Acids Res. 2008 Jan;36(Database issue):D178-83
references.pmid: 17947322
references.citation: Nucleic Acids Res. 2013 Jan;41(Database issue):D262-7
references.pmid: 23118484
references.citation: Gene. 1987;58(2-3):201-16
references.pmid: 2828185
references.citation: RNA. 2010 Jul;16(7):1317-27
references.pmid: 20484468
references.citation: Biochemistry. 1999 Oct 5;38(40):13338-46
references.pmid: 10529209
references.citation: Nature. 2003 Oct 16;425(6959):686-91
references.pmid: 14562095
references.citation: Free Radic Biol Med. 2011 Oct 15;51(8):1589-600
references.pmid: 21821119
references.citation: Mol Cell Biol. 2008 May;28(10):3151-61
references.pmid: 18332120
references.citation: Nat Protoc. 2009;4(3):363-71
references.pmid: 19247286
references.citation: Nat Struct Mol Biol. 2006 Jan;13(1):13-21
references.pmid: 16369484
references.citation: EMBO Rep. 2007 Aug;8(8):763-9
references.pmid: 17599088
references.citation: RNA. 2008 Jan;14(1):98-106
references.pmid: 18025251
references.citation: Bioinformatics. 2000 Apr;16(4):404-5
references.pmid: 10869041
references.citation: Trends Biochem Sci. 2002 Jul;27(7):344-51
references.pmid: 12114023
references.citation: PLoS Genet. 2012;8(6):e1002732
references.pmid: 22685417
references.citation: Nucleic Acids Res. 1999 Sep 1;27(17):3543-9
references.pmid: 10446245
references.citation: Mol Cell. 2007 Dec 28;28(6):965-77
references.pmid: 18158895
references.citation: Biochemistry. 1994 Nov 15;33(45):13475-83
references.pmid: 7947756
references.citation: Mol Cell Biol. 2012 Jun;32(12):2254-67
references.pmid: 22493060
references.citation: J Chromatogr. 1989 Jun 2;471:3-36
references.pmid: 2670985
references.citation: PLoS One. 2007 Jan 24;2(1):e174
references.pmid: 17245450
references.citation: Mol Cell Biol. 2005 Dec;25(23):10352-64
references.pmid: 16287850
references.citation: Mol Microbiol. 2000 Feb;35(4):936-48
references.pmid: 10692169
references.citation: Eur J Biochem. 1973 Nov 15;39(2):325-33
references.pmid: 4775053
references.citation: Nucleic Acids Res. 2002 Oct 15;30(20):4509-18
references.pmid: 12384598
references.citation: Eur J Biochem. 1977 May 2;75(1):311-8
references.pmid: 405217
references.citation: Trends Biochem Sci. 2003 Jun;28(6):329-35
references.pmid: 12826405
references.citation: Curr Opin Struct Biol. 2008 Jun;18(3):330-9
references.pmid: 18539024
references.citation: Wiley Interdiscip Rev RNA. 2011 Sep-Oct;2(5):611-31
references.pmid: 21823225
references.citation: Biochemistry. 2012 Jun 26;51(25):5091-104
references.pmid: 22650761
references.citation: Nucleic Acids Res. 2001 Oct 1;29(19):3997-4005
references.pmid: 11574682
references.citation: Nucleic Acids Res. 2007;35(12):4018-29
references.pmid: 17553838
references.citation: Nucleic Acids Res. 2011 Mar;39(4):1526-37
references.pmid: 20972225
references.citation: Proteins. 2008 Feb 15;70(3):611-25
references.pmid: 17876813
references.citation: Bioessays. 2012 Sep;34(9):725-9
references.pmid: 22706794
references.citation: Mol Cell. 2004 Nov 19;16(4):663-9
references.pmid: 15546625
references.citation: Methods Enzymol. 1983;101:228-45
references.pmid: 6310326
references.citation: Chembiochem. 2001 Jan 8;2(1):31-5
references.pmid: 11828423
references.citation: EMBO J. 1994 Feb 15;13(4):906-13
references.pmid: 8112304
references.citation: Bioinformatics. 2004 Sep 1;20(13):2138-9
references.pmid: 15044227
references.citation: Science. 2000 Aug 11;289(5481):920-30
references.pmid: 10937990
references.citation: RNA. 2004 Nov;10(11):1713-20
references.pmid: 15388872
references.citation: Aging (Albany NY). 2009 Dec 04;1(11):957-60
references.pmid: 20157578
references.citation: Nucleic Acids Res. 2013 Jan;41(2):1151-63
references.pmid: 23180764
references.citation: Anal Biochem. 1989 Feb 1;176(2):303-6
references.pmid: 2662810
references.citation: Cell. 2012 Jun 22;149(7):1635-46
references.pmid: 22608085
references.citation: J Biol Chem. 2008 Sep 26;283(39):26548-56
references.pmid: 18667428
references.citation: PLoS Genet. 2010 Dec 16;6(12):e1001247
references.pmid: 21187895
references.citation: Nucleic Acids Res. 2012 Dec;40(22):11648-58
references.pmid: 23074192
references.citation: RNA. 2004 Jun;10(6):907-13
references.pmid: 15146074
references.citation: Nucleic Acids Res. 2010 Sep;38(16):5327-35
references.pmid: 20421205
references.citation: J Mol Biol. 1994 Aug 19;241(3):492-7
references.pmid: 8064863
references.citation: PLoS One. 2011;6(8):e23168
references.pmid: 21858014
references.citation: Nucleic Acids Res. 2008 Apr;36(6):1826-35
references.pmid: 18263608
references.citation: Proteins. 2002 May 1;47(2):228-35
references.pmid: 11933069
references.citation: RNA. 2008 Oct;14(10):2223-33
references.pmid: 18755836
references.citation: Nucleic Acids Res. 1981 Dec 21;9(24):6935-52
references.pmid: 7335496
references.citation: Acc Chem Res. 2011 Dec 20;44(12):1380-8
references.pmid: 21615108