Protein synthesis in Escherichia coli with mischarged tRNA |
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| Authors: | Min Bokkee Kitabatake Makoto Polycarpo Carla Pelaschier Joanne Raczniak Gregory Ruan Benfang Kobayashi Hiroyuki Namgoong Suk Söll Dieter |
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| Affiliation: | Department of Molecular Biophysics, Yale University, New Haven, Connecticut 06520-8114, USA. |
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| Abstract: | Two types of aspartyl-tRNA synthetase exist: the discriminating enzyme (D-AspRS) forms only Asp-tRNA(Asp), while the nondiscriminating one (ND-AspRS) also synthesizes Asp-tRNA(Asn), a required intermediate in protein synthesis in many organisms (but not in Escherichia coli). On the basis of the E. coli trpA34 missense mutant transformed with heterologous ND-aspS genes, we developed a system with which to measure the in vivo formation of Asp-tRNA(Asn) and its acceptance by elongation factor EF-Tu. While large amounts of Asp-tRNA(Asn) are detrimental to E. coli, smaller amounts support protein synthesis and allow the formation of up to 38% of the wild-type level of missense-suppressed tryptophan synthetase. |
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