PMID:2085546

Citation	Yancey, SD and Kushner, SR (1990) Isolation and characterization of a new temperature-sensitive polynucleotide phosphorylase mutation in Escherichia coli K-12. Biochimie 72:835-43
Abstract	Polynucleotide phosphorylase (PNPase) has been studied in detail since its discovery in 1955 [1]. In an attempt to determine what role, if any, it has in mRNA decay in Escherichia coli, we have isolated and characterized a temperature-sensitive mutation, pnp-200, in the pnp gene. In vitro phosphorolysis, polymerization and exchange activities of the partially purified Pnp-200 enzyme are all reduced to 30-40% of wild-type activity at 50 degrees C compared to 32 degrees C. The pnp-200 mutation alone does not affect cell growth or mRNA stability. A triple mutant strain containing pnp-200 in combination with other temperature-sensitive mutations in genes known to affect mRNA metabolism (rnb-500 and ams-1) is conditionally lethal and shows increased mRNA stability after shift to the non-permissive temperature.
Links	PubMed
Keywords	Escherichia coli/enzymology; Escherichia coli/genetics; Genes, Bacterial; Genotype; Hot Temperature; Kinetics; Mutation; Plasmids; Polyribonucleotide Nucleotidyltransferase/genetics; Polyribonucleotide Nucleotidyltransferase/isolation & purification; Polyribonucleotide Nucleotidyltransferase/metabolism; Restriction Mapping; Thermodynamics
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