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Misacylation of pyrrolysine tRNA in vitro and in vivo
Author(s) -
Gundllapalli Sarath,
Ambrogelly Alexandre,
Umehara Takuya,
Li Darrick,
Polycarpo Carla,
Söll Dieter
Publication year - 2008
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2008.08.027
Subject(s) - methanosarcina barkeri , aminoacylation , transfer rna , escherichia coli , biology , biochemistry , serine , in vitro , microbiology and biotechnology , gene , enzyme , genetics , rna , bacteria , methanogenesis
Methanosarcina barkeri inserts pyrrolysine (Pyl) at an in‐frame UAG codon in its monomethylamine methyltransferase gene. Pyrrolysyl‐tRNA synthetase acylates Pyl onto tRNA Pyl , the amber suppressor pyrrolysine Pyl tRNA. Here we show that M. barkeri Fusaro tRNA Pyl 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 tRNA Pyl contains two base changes; a G3:U70 pair, the known identity element for E. coli alanyl‐tRNA synthetase (AlaRS). While M. barkeri MS tRNA Pyl cannot be alanylated by E. coli AlaRS, mutation of the MS tRNA Pyl A4:U69 pair into C4:G69 allows aminoacylation by E. coli AlaRS both in vitro and in vivo.