Summary
Eight suppressors of trpA218, a missense double mutant of trpA, the gene for the tryptophan synthetase alpha chain of Escherichia coli, have been further characterized genetically, physiologically and biochemically. trpA218 possesses an inactive alpha chain that contains leucine (instead of phenylalanine) at position 22 and serine (instead of glycine) at position 211. Replacement of either mutant amino acid by the corresponding wild type amino acid leads to an active alpha chain. To determine whether each trpA218 suppressor (Su218) affects the 22 or 211 position, a “substitute” trpA218 was constructed. Whereas the original double mutant possesses a Ser211 specified by the codon AGU, we constructed a trpA(Leu22-Ser211) in which the Ser211 codon is UCG. All eight Su218s failed to suppress the new double mutant. The suppressors fall into two classes according to growth in various media. Six of the eight map in the region of glyV, a gene for the GGU/C-reading glycine tRNA. After reversed phase column chromatography of radioactively labeled glycyl-tRNA, the suppressor tRNAs exhibited altered profiles that were similarly different from the parental tRNA in all eight cases. These results suggest that there are several classes of Su218, that all of them suppress the serine codon AGU (or AGC) corresponding to position 211, and that at least six of the eight are mutationally altered glycine tRNAs.
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Allen, M.K., Yanfsky, C.: A biochemical and genetic study of reversion with the A-gene A-protein system of Escherichia coli tryptophan synthetase. Genetics 48, 1065–1083 (1963)
Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K-12. Bact. Rev. 40, 116–167 (1976)
Carbon, J., Curry, J.B.: Genetically and chemically derived missense suppressor transfer RNA's with altered enzymic aminoacylation rates. J. molec. Biol. 38, 201–216 (1968)
Carbon, J., Fleck, E.W.: Genetic alteration of structure and function in glycine transfer RNA of Escherichia coli: mechanism of suppression of the tryptophan synthetase A78 mutation. J. molec. Biol. 85, 371–391 (1974)
Fleck, E.W., Carbon, J.: Multiple gene loci for a single species of glycine transfer ribonucleic acid. J. Bact. 122, 492–501 (1975)
Fredericq, P.: The recombination of colicinogenic factors with other episomes and plasmids, pp. 163–174. In: Bacterial plasmids and episomes (G.E.W. Wolstenholme and M. O'Connor, eds.). Boston: Little, Brown and Co., 1969
Hill, C.W., Combriato, G., Steinhart, W., Riddle, D.L., Carbon, J.: The nucleotide sequence of the GGG-specific glycine transfer ribonucleic acid of Escherichia coli and of Salmonella typhimurium. J. biol. Chem. 248, 4252–4262 (1973)
Ilgen, C., Kirk, L.L., Carbon, J.: The isolation and characterization of large transfer ribonucleic acid precursors from Escherichia coli. J. biol. Chem. 251, 922–929 (1976)
Murgola, E.J., Prather, N.E., Hadley, K.H.: Variations among glyV-derived glycine tRNA suppressors of glutamic acid codons. J. Bact. 134, 801–807 (1978)
Murgola, E.J., Yanofsky, C.: Structural interactions between amino acid residues at positions 22 and 211 in the tryptophan synthetase alpha chain of Escherichia coli. J. Bact. 117, 444–448 (1974a)
Murgola, E.J., Yanofsky, C.: Selection for new amino acids at position 211 of the tryptophan synthetase alpha chain of Escherichia coli. J. molec. Biol. 86, 775–784 (1974b)
Murgola, E.J., Yanofsky, C.: Suppression of glutamic acid codons by mutant glycine transfer ribonucleic acid. J. Bact. 117, 439–443 (1974c)
Ostrem, D.L., Berg, P.: Glycyl transfer ribonucleic acid synthetase from Escherichia coli; purification, properties and substrate binding. Biochemistry 13, 1338–1348 (1974)
Pearson, R.L., Weiss, J.F., Kelmers, A.D.: Improved separation of transfer RNAs on polychlorotrifluoroethylene-supported reversed phase chromatography columns. Biochim. biophys. Acta (Amst.) 228, 770–774 (1971)
Roberts, J.W., Carbon, J.: Molecular mechanism for missense suppression in E. coli. Nature (Lond.) 250, 412–414 (1974)
Roberts, J.W., Carbon, J.: Nucleotide sequence studies of normal and genetically altered glycine transfer ribonucleic acids from Escherichia coli. J. biol. Chem. 250, 5530–5541 (1975)
Squires, C., Carbon, J.: Normal and mutant glycine transfer RNAs. Nature (Lond.) New Biol. 233, 274–277 (1971)
Yanofsky, C.: Protein structure and evolution. In M. Marois (ed.), Proc. 2nd Int. Conf. on Theoret. Phys. Biol., Centre National de la Recherche Scientifique, Paris, pp. 191–206 (1969)
Yanofsky, C., Horn, V.: Tryptophan synthetase alpha chain positions affected by mutations near the ends of the genetic map of trpA of Escherichia coli. J. biol. Chem. 247, 4494–4498 (1972)
Yanofsky, C., Ito, J., Horn, V.: Amino acid replacements and the genetic code. Cold Spr. Harb. Symp. quant. Biol. 31, 151–162 (1966)
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Murgola, E.J., Mims, B.H. & Prather, N.E. Characterization of missense suppressors of a double mutant of the tryptophan synthetase alpha chain of Escherichia coli . Molec. Gen. Genet. 165, 225–230 (1978). https://doi.org/10.1007/BF00269911
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DOI: https://doi.org/10.1007/BF00269911