title: Genetic properties of temperature-sensitive folding mutants of the coat protein of phage P22.
abstract: Temperature-sensitive mutations fall into two general classes: those generating thermolabile proteins; and those generating defects in protein synthesis, folding or assembly. Temperature-sensitive mutations at 17 sites in the gene for the coat protein of Phage P22 are of the latter class, preventing the productive folding of the polypeptide chain at restrictive temperature. We show here that, though the coat subunits interact intimately to form the viral shell, these temperature-sensitive folding (TSF) mutations were all recessive to wild type. The mutant polypeptide chains were not rescued by the presence of wild-type polypeptide chains. Missense substitutions in multimeric proteins frequently exhibit intragenic complementation; however, all pairs of coat protein TSF mutants tested failed to complement. The recessive phenotypes, absence of rescue and absence of intragenic complementation are all accounted for by the TSF defect, in which destabilization of a folding intermediate at restrictive temperature prevents the mutant chain from reaching the conformation required for subunit/subunit recognition. We suggest that absence of intragenic complementation should be a general property of TSF mutations in genes encoding multimeric proteins. The spectra of new loci identified by isolating second-site suppressors and synthetic lethals of temperature sensitive mutants will also differ depending on the nature of the defect. In the case of TSF mutations, where folding intermediates are defective rather than the native molecule, the spectra of other genes identified should shift from those whose products interact with the native molecule to those whose products influence the folding process.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
authors.forename: C L
authors.initials: CL
authors.lastname: Gordon
authors.forename: J
authors.initials: J
authors.lastname: King
mesh_terms: D017100:Bacteriophage P22 / Q000235:genetics*; D002213:Capsid / Q000737:chemistry / Q000235:genetics*; D036022:Capsid Proteins; D005808:Genes, Recessive; D016153:Genes, Suppressor; D005814:Genes, Viral; D005816:Genetic Complementation Test; D016296:Mutagenesis; D010641:Phenotype; D011489:Protein Denaturation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D013696:Temperature; D015678:Viral Structural Proteins / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D015678:Viral Structural Proteins; C079841:portal protein, bacteriophage P22