Elongator function in tRNA wobble uridine modification is conserved between yeast and plants.
Based on studies in yeast and mammalian cells the Elongator complex has been implicated in functions as diverse as histone acetylation, polarized protein trafficking and tRNA modification. Here we show that Arabidopsis mutants lacking the Elongator subunit AtELP3/ELO3 have a defect in tRNA wobble uridine modification. Moreover, we demonstrate that yeast elp3 and elp1 mutants expressing the respective Arabidopsis Elongator homologues AtELP3/ELO3 and AtELP1/ELO2 assemble integer Elongator complexes indicating a high degree of structural conservation. Surprisingly, in vivo complementation studies based on Elongator-dependent tRNA nonsense suppression and zymocin tRNase toxin assays indicated that while AtELP1 rescued defects of a yeast elp1 mutant, the most conserved Elongator gene AtELP3, failed to complement an elp3 mutant. This lack of complementation is due to incompatibility with yeast ELP1 as coexpression of both plant genes in an elp1 elp3 yeast mutant restored Elongator's tRNA modification function in vivo. Similarly, AtELP1, not ScELP1 also supported partial complementation by yeast-plant Elp3 hybrids suggesting that AtElp1 has less stringent sequence requirements for Elp3 than ScElp1. We conclude that yeast and plant Elongator share tRNA modification roles and propose that this function might be conserved in Elongator from all eukaryotic kingdoms of life.
Mehlgarten
Constance
C
Institut für Biologie, Genetik, Martin-Luther-Universität Halle-Wittenberg, Weinbergweg 10, D-06120 Halle (Saale), Germany.
Jablonowski
Daniel
D
Wrackmeyer
Uta
U
Tschitschmann
Susan
S
Sondermann
David
D
Jäger
Gunilla
G
Gong
Zhizhong
Z
Byström
Anders S
AS
Schaffrath
Raffael
R
Breunig
Karin D
KD
D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D015394:Molecular Structure; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014529:Uridine / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D029681:Arabidopsis Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer; D000123:Acetyltransferases; C107118:ELO2 protein, S cerevisiae; C502551:ELO3 protein, Arabidopsis; C526363:Elp3 protein, S cerevisiae; D051548:Histone Acetyltransferases; D014529:Uridine
Gene. 1995 Apr 14;156(1):119-22
7737504
Biosci Biotechnol Biochem. 1996 May;60(5):798-801
8704309
Cell. 2009 Feb 6;136(3):551-64
19185337
Mol Microbiol. 2002 Aug;45(3):817-26
12139626
Science. 1996 Nov 15;274(5290):1201-4
8895471
Proc Natl Acad Sci U S A. 1985 Aug;82(15):4905-9
3860833
Cell. 2007 Nov 16;131(4):633-6
18022353
Mol Cell Biol. 2006 Sep;26(18):6902-12
16943431
EMBO J. 2004 Oct 27;23(21):4243-52
15457216
J Chromatogr. 1982 Jul 9;230(2):297-308
7050138
Mol Microbiol. 2002 Feb;43(3):783-91
11929532
Trends Biochem Sci. 2002 Mar;27(3):115-7
11893502
Yeast. 2001 Oct;18(14):1285-99
11571753
Mol Cell Biol. 2001 Dec;21(23):8203-12
11689709
Am J Hum Genet. 2001 Mar;68(3):753-8
11179021
Mol Cell. 2002 Oct;10(4):925-33
12419235
Proc Natl Acad Sci U S A. 2009 Sep 15;106(37):15645-50
19805217
Mol Microbiol. 2006 Feb;59(3):795-806
16420352
Mol Cell. 1999 Jul;4(1):123-8
10445034
J Biol Chem. 2005 May 20;280(20):19454-60
15772087
Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1678-83
20080602
Anal Biochem. 1976 May 7;72:248-54
942051
Mol Cell. 2006 May 19;22(4):521-31
16713582
Curr Opin Cell Biol. 2007 Jun;19(3):331-6
17466506
Mol Cell. 2004 May 21;14(4):457-64
15149595
Nucleic Acids Res. 2008 Mar;36(4):1072-80
18096622
PLoS Genet. 2010 Jan 22;6(1):e1000820
20107598
Yeast. 2004 Aug;21(11):947-62
15334558
Eukaryot Cell. 2009 Sep;8(9):1362-72
19633267
Mol Cell. 2007 Dec 14;28(5):860-70
18082610
Mol Microbiol. 2008 Sep;69(5):1266-77
18657261
Plant Cell. 2003 Mar;15(3):639-54
12615938
Mol Microbiol. 2006 Jan;59(2):677-88
16390459
Mol Microbiol. 2003 Sep;49(5):1297-307
12940988
Mol Microbiol. 2009 Sep;73(5):869-81
19656297
Curr Genet. 1989 Dec;16(5-6):339-46
2692852
RNA. 2005 Apr;11(4):424-36
15769872
Mol Genet Genomics. 2005 May;273(3):264-72
15902492
Yeast. 1999 Jul;15(10B):963-72
10407276
Mol Cell. 2005 Mar 18;17(6):841-53
15780940
Nature. 2010 Jan 28;463(7280):554-8
20054296
Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3517-22
11904415
J Mol Biol. 1994 Jul 1;240(1):8-19
8021943
Mol Cell. 2006 Oct 6;24(1):139-48
17018299
EMBO J. 2001 Jan 15;20(1-2):231-9
11226173
EMBO J. 2001 Apr 17;20(8):1993-2003
11296232
Fungal Genet Biol. 2000 Aug;30(3):173-90
11035939
Proc Natl Acad Sci U S A. 2005 May 24;102(21):7754-9
15894610
Methods Enzymol. 1991;194:1-863
2005781
Biochemistry. 2005 Jun 7;44(22):8078-89
15924427
RNA. 2005 Nov;11(11):1648-54
16244131
Am J Hum Genet. 2001 Mar;68(3):598-605
11179008
PLoS Genet. 2009 Jul;5(7):e1000561
19593383
Mol Cell. 1999 Jan;3(1):109-18
10024884
Mol Cell Biol. 2008 May;28(10):3301-12
18332122
J Biol Chem. 2003 Jan 10;278(2):956-61
12424236
J Biol Chem. 2004 Jul 30;279(31):32087-92
15138274
PLoS Genet. 2009 Oct;5(10):e1000684
19834596