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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Alanine / chemistry
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Alanine / metabolism
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Alanine-tRNA Ligase / chemistry
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Alanine-tRNA Ligase / metabolism
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Archaeal Proteins / genetics
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Archaeal Proteins / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Lysine / analogs & derivatives*
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Lysine / chemistry
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Lysine / metabolism
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Methanosarcina barkeri / enzymology
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Methanosarcina barkeri / genetics
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Methanosarcina barkeri / metabolism*
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Methyltransferases / genetics
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Methyltransferases / metabolism
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Mutation
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RNA, Archaeal / chemistry
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RNA, Archaeal / metabolism*
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RNA, Transfer, Lys / chemistry
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RNA, Transfer, Lys / metabolism*
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Serine / chemistry
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Serine / metabolism
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Serine-tRNA Ligase / chemistry
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Serine-tRNA Ligase / metabolism
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Transfer RNA Aminoacylation*
Substances
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Archaeal Proteins
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RNA, Archaeal
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RNA, Transfer, Lys
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Serine
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Methyltransferases
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monomethylamine methyltransferase
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Serine-tRNA Ligase
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Alanine-tRNA Ligase
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pyrrolysine
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Lysine
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Alanine