Mistranslation in bacteriophage-infected anucleate minicells of Escherichia coli: a test for error propagation

The theory of error propagation proposes that errors occurring during expression of the genetic code lead to increased levels of error occurrence in successive generations. A model system for testing error propagation in bacteriophage T7 infected anucleate minicells of Escherichia coli is described. Errors in translation were were stimulated by addition of gentamicin to phage infected minicells, and the error frequency based on the illegitimate incorporation of L-[35S] cysteine into the T7 0.3 gene protein calculated to be on the order of 1 error per 10 000 codons translated. Errors in the synthesis of T7 early gene products have also been detected as suppression of a UAG nonsense codon in gene 1 of the T7 DNA-dependent RNA polymerase, and as increased charge heterogeneity in the gene 1.3 product (DNA ligase). The question of error propagation has been addressed by infecting minicells with a mutant of T7 containing nonsense mutations in the early gene 1 and late gene 16. Results demonstrate that a T7 DNA-dependent RNA polymerase containing misincorporated amino acids is unable, by mistranscription, to suppress a UAG nonsense codon located in the late T7 gene 16.