title: Misacylation of pyrrolysine tRNA in vitro and in vivo.
abstract: 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.
authors.affiliation: Departments of Molecular Biophysics and Biochemistry, Yale University, P.O. Box 208114, 266 Whitney Avenue, New Haven, CT 06520-8114, USA.
authors.forename: Sarath
authors.initials: S
authors.lastname: Gundllapalli
authors.forename: Alexandre
authors.initials: A
authors.lastname: Ambrogelly
authors.forename: Takuya
authors.initials: T
authors.lastname: Umehara
authors.forename: Darrick
authors.initials: D
authors.lastname: Li
authors.forename: Carla
authors.initials: C
authors.lastname: Polycarpo
authors.forename: Dieter
authors.initials: D
authors.lastname: Söll
mesh_terms: 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
chemical_list: 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
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