Misacylation of pyrrolysine tRNA in vitro and in vivo.
Methanosarcina barkeri inserts pyrrolysine (Pyl) at an in-frame UAG codon in its monomethylamine methyltransferase gene. Pyrrolysyl-tRNA synthetase acylates Pyl onto tRNAPyl, the amber suppressor pyrrolysine Pyl tRNA. Here we show that M. barkeri Fusaro tRNAPyl can be misacylated with serine by the M. barkeri bacterial-type seryl-tRNA synthetase in vitro and in vivo in Escherichia coli. Compared to the M. barkeri Fusaro tRNA, the M. barkeri MS tRNAPyl contains two base changes; a G3:U70 pair, the known identity element for E. coli alanyl-tRNA synthetase (AlaRS). While M. barkeri MS tRNAPyl cannot be alanylated by E. coli AlaRS, mutation of the MS tRNAPyl A4:U69 pair into C4:G69 allows aminoacylation by E. coli AlaRS both in vitro and in vivo.
Gundllapalli
Sarath
S
Departments of Molecular Biophysics and Biochemistry, Yale University, P.O. Box 208114, 266 Whitney Avenue, New Haven, CT 06520-8114, USA.
Ambrogelly
Alexandre
A
Umehara
Takuya
T
Li
Darrick
D
Polycarpo
Carla
C
Söll
Dieter
D
D000409:Alanine / Q000737:chemistry / Q000378:metabolism; D000412:Alanine-tRNA Ligase / Q000737:chemistry / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D017021:Methanosarcina barkeri / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D019642:RNA, Archaeal / Q000737:chemistry / Q000378:metabolism*; D012357:RNA, Transfer, Lys / Q000737:chemistry / Q000378:metabolism*; D012694:Serine / Q000737:chemistry / Q000378:metabolism; D012714:Serine-tRNA Ligase / Q000737:chemistry / Q000378:metabolism; D046249:Transfer RNA Aminoacylation
D019843:Archaeal Proteins; D019642:RNA, Archaeal; D012357:RNA, Transfer, Lys; D012694:Serine; D008780:Methyltransferases; C113200:monomethylamine methyltransferase; D012714:Serine-tRNA Ligase; D000412:Alanine-tRNA Ligase; C456839:pyrrolysine; D008239:Lysine; D000409:Alanine
Nucleic Acids Res. 2007;35(4):1270-8
17267409
Nat Chem Biol. 2007 Jan;3(1):29-35
17173027
Nat Chem Biol. 2008 Apr;4(4):232-4
18278036
Proc Natl Acad Sci U S A. 2004 Aug 24;101(34):12450-4
15314242
Science. 1988 May 6;240(4853):793-6
2452483
Nature. 1988 May 12;333(6169):140-5
3285220
J Mol Biol. 2008 May 2;378(3):634-52
18387634
Proc Natl Acad Sci U S A. 2007 Jul 3;104(27):11268-73
17592110
J Bacteriol. 1998 Dec;180(24):6446-9
9851985
Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3141-6
17360621
J Biol Chem. 2004 Nov 19;279(47):48780-6
15364939
Biochem Biophys Res Commun. 2008 Sep 26;374(3):470-4
18656445
Nucleic Acids Res. 1994 Dec 11;22(24):5378-84
7529407
Science. 2002 May 24;296(5572):1459-62
12029131
Trends Biochem Sci. 1989 Jun;14(6):233-7
2669241
Curr Opin Microbiol. 2005 Dec;8(6):706-12
16256420
FEBS Lett. 2007 Jul 10;581(17):3197-203
17582401
Annu Rev Genet. 1985;19:57-80
2417544
Mol Cell. 2004 Jan 30;13(2):157-68
14759362
Mol Cell. 2003 Aug;12(2):287-94
14536069
Nature. 2004 Sep 16;431(7006):333-5
15329732
Nucleic Acids Res. 2004 Feb 10;32(3):1091-6
14872064
EMBO J. 2005 Oct 5;24(19):3369-79
16163389
Biochem Biophys Res Commun. 2008 Jul 11;371(4):818-22
18471995
FEBS Lett. 2007 Nov 13;581(27):5282-8
17967457
FEBS Lett. 2006 Dec 11;580(28-29):6695-700
17126325