title: Translational misreading in Mycobacterium smegmatis increases in stationary phase.
abstract: The study of errors in gene translation has largely been confined to a small number of model organisms. We have examined all possible misreading errors at a defined codon in Mycobacterium smegmatis. Using a dual-luciferase gain of function reporter system that employs a mutated essential lysine in firefly luciferase, we accurately quantified mistranslation errors. Overall, accuracy of gene translation was comparable with Escherichia coli at <1/2000 errors/codon during exponential growth. Stationary phase was associated with a dramatic increase in misincorporation errors by Lys-tRNACUU(Lys) at a subset of three codons, each with a single base changed from the AAG lysine codon. The maximum error rate detected was 0.2% with codon AUG. Treatment with streptomycin increased misreading errors at several codons associated in particular with U·U, G·U and C·U codon·anti-codon mismatches, but oxidative stress did not change translational fidelity. Our study is the first comprehensive examination of misreading errors for a defined codon in mycobacteria.
authors.affiliation: Collaborative Innovation Centre for the Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University, Beijing, 100084, China.
authors.forename: Tianqi
authors.initials: T
authors.lastname: Leng
authors.affiliation: Collaborative Innovation Centre for the Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University, Beijing, 100084, China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Pan
authors.affiliation: Collaborative Innovation Centre for the Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University, Beijing, 100084, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Xu
authors.affiliation: Collaborative Innovation Centre for the Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University, Beijing, 100084, China. Electronic address: bjavid@gmail.com.
authors.forename: Babak
authors.initials: B
authors.lastname: Javid
mesh_terms: D003062:Codon; D048168:Escherichia coli K12 / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D005838:Genotype; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics*; D009154:Mutation; D020102:Mycobacterium smegmatis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D018384:Oxidative Stress; D014176:Protein Biosynthesis / Q000187:drug effects; D012357:RNA, Transfer, Lys / Q000235:genetics / Q000378:metabolism; D013307:Streptomycin / Q000494:pharmacology
chemical_list: D003062:Codon; D012357:RNA, Transfer, Lys; D049409:Luciferases, Firefly; D013307:Streptomycin
keywords: Decoding errors; Mistranslation; Mycobacterium