Adding pyrrolysine to the Escherichia coli genetic code.
Pyrrolysyl-tRNA synthetase and its cognate suppressor tRNA(Pyl) mediate pyrrolysine (Pyl) insertion at in frame UAG codons. The presence of an RNA hairpin structure named Pyl insertion structure (PYLIS) downstream of the suppression site has been shown to stimulate the insertion of Pyl in archaea. We study here the impact of the presence of PYLIS on the level of Pyl and the Pyl analog N-epsilon-cyclopentyloxycarbonyl-l-lysine (Cyc) incorporation using a quantitative lacZ-luc tandem reporter system in an Escherichia coli context. We show that PYLIS has no effect on the level of neither Pyl nor Cyc incorporation. Exogenously supplying our reporter system with d-ornithine significantly increases suppression efficiency, indicating that d-ornithine is a direct precursor to Pyl.
Namy
Olivier
O
Institut de Genetique et Microbiologie, Université Paris-Sud, CNRS UMR8621, Orsay F-91405, France.
Zhou
Yu
Y
Gundllapalli
Sarath
S
Polycarpo
Carla R
CR
Denise
Alain
A
Rousset
Jean-Pierre
JP
Söll
Dieter
D
Ambrogelly
Alexandre
A
D019843:Archaeal Proteins / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D019766:Genes, Archaeal; D005798:Genes, Bacterial; D005815:Genetic Code; D008239:Lysine / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D017021:Methanosarcina barkeri / Q000235:genetics; D008780:Methyltransferases / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009952:Ornithine / Q000378:metabolism; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D019843:Archaeal Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D009952:Ornithine; D008780:Methyltransferases; C113200:monomethylamine methyltransferase; C456839:pyrrolysine; D008239:Lysine