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PMID: 2187197 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Escherichia coli thymidylate synthase: amino acid substitutions by suppression of amber nonsense mutations.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 10 ·1990-05-00 ·Pages 3957-61

Michaels ML, Kim CW, Matthews DA, Miller JH

Abstract

By using site-directed oligonucleotide mutagenesis, amber nonsense stop codons (5'-TAG-3') have been introduced at 20 sites in the Escherichia coli thymidylate synthase gene. By transforming the thyA mutant plasmids into 13 strains, each of which harbor different amber suppressor tRNAs, we were able to generate over 245 amino acid substitutions in E. coli thymidylate synthase (EC 2.1.1.45). Growth characteristics of these mutants have been studied, yielding a body of information that includes some surprising results in light of the recently published crystal structure of the enzyme.

MeSH Terms
Amino Acid Sequence Amino Acids Escherichia coli/enzymology,genetics,growth & development Genes, Bacterial Models, Molecular Mutation Plasmids Protein Conformation Restriction Mapping Suppression, Genetic Thymidylate Synthase/genetics,metabolism
Chemicals
Amino Acids Thymidylate Synthase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Michaels M L
Molecular Biology Institute, University of California, Los Angeles 90024.
Kim C W
Matthews D A
Miller J H
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.10.3957
Published
1990-05-00
Pages
3957-61
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC54023
Subset
IM
Grants
NIGMS NIH HHS · GM-07104 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.10.3957 PMC Full Text View journal details
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