Measurements of translation initiation from all 64 codons in E. coli.
Our understanding of translation underpins our capacity to engineer living systems. The canonical start codon (AUG) and a few near-cognates (GUG, UUG) are considered as the 'start codons' for translation initiation in Escherichia coli. Translation is typically not thought to initiate from the 61 remaining codons. Here, we quantified translation initiation of green fluorescent protein and nanoluciferase in E. coli from all 64 triplet codons and across a range of DNA copy number. We detected initiation of protein synthesis above measurement background for 47 codons. Translation from non-canonical start codons ranged from 0.007 to 3% relative to translation from AUG. Translation from 17 non-AUG codons exceeded the highest reported rates of non-cognate codon recognition. Translation initiation from non-canonical start codons may contribute to the synthesis of peptides in both natural and synthetic biological systems.
Joint Initiative for Metrology in Biology, Stanford, CA 94305, USA.|Genome-scale Measurements Group, National Institute of Standards and Technology, Stanford, CA 94305, USA.|Department of Bioengineering, Stanford, CA 94305, USA.
Ariel
A
Hecht
Joint Initiative for Metrology in Biology, Stanford, CA 94305, USA.|Genome-scale Measurements Group, National Institute of Standards and Technology, Stanford, CA 94305, USA.|Department of Bioengineering, Stanford, CA 94305, USA.
Jeff
J
Glasgow
Department of Bioengineering, Stanford, CA 94305, USA.|Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW 2109, Australia.
Paul R
PR
Jaschke
Joint Initiative for Metrology in Biology, Stanford, CA 94305, USA.|Genome-scale Measurements Group, National Institute of Standards and Technology, Stanford, CA 94305, USA.|Department of Bioengineering, Stanford, CA 94305, USA.
Lukmaan A
LA
Bawazer
Joint Initiative for Metrology in Biology, Stanford, CA 94305, USA.|Genome-scale Measurements Group, National Institute of Standards and Technology, Stanford, CA 94305, USA.|Department of Bioengineering, Stanford, CA 94305, USA.
Matthew S
MS
Munson
Joint Initiative for Metrology in Biology, Stanford, CA 94305, USA.|Department of Bioengineering, Stanford, CA 94305, USA.
Jennifer R
JR
Cochran
Joint Initiative for Metrology in Biology, Stanford, CA 94305, USA.|Department of Bioengineering, Stanford, CA 94305, USA.
Drew
D
Endy
Joint Initiative for Metrology in Biology, Stanford, CA 94305, USA.|Genome-scale Measurements Group, National Institute of Standards and Technology, Stanford, CA 94305, USA.|Department of Bioengineering, Stanford, CA 94305, USA.
Marc
M
Salit
D003062:Codon; D018387:Codon, Initiator; D004926:Escherichia coli / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D010442:Peptide Chain Initiation, Translational; D010957:Plasmids / Q000235:genetics
D003062:Codon; D018387:Codon, Initiator; D049452:Green Fluorescent Proteins; D008156:Luciferases