Intragenic suppressors of folding defects in the P22 tailspike protein

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Within the amino acid sequences of polypeptide chains little is known of the distribution of sites and sequences critical for directing chain folding and assembly. Temperature-sensitive folding (tsf) mutations identifying such sites have been previously isolated and characterized in gene 9 of phage P22 encoding the tailspike endorhamnosidase. We report here the isolation of a set of second-site conformational suppressors which alleviate the defect in such folding mutants. The suppressors were selected for their ability to correct the defects of missense tailspike polypeptide chains, generated by growth of gene 9 amber mutants on Salmonella host strains inserting either tyrosine, serine, glutamine or leucine at the nonsense codons. Second-site suppressors were recovered for 13 of 22 starting sites. The suppressors of defects at six sites mapped within gene 9. (Suppressors for seven other sites were extragenic and distant from gene 9.) The missense polypeptide chains generated from all six suppressible sites displayed ts phenotypes. Temperature-sensitive alleles were isolated at these amber sites by pseudoreversion. The intragenic suppressors restored growth at the restrictive temperature of these presumptive tsf alleles. Characterization of protein maturation in cells infected with mutant phages carrying the intragenic suppressors indicates that the suppression is acting at the level of polypeptide chain folding and assembly.

Original languageEnglish (US)
Pages (from-to)263-277
Number of pages15
JournalGenetics
Volume127
Issue number2
StatePublished - 1991
Externally publishedYes

Fingerprint

Amber
Peptides
Temperature
Genes
Proteins
Bacteriophage P22
Alleles
Nonsense Codon
Growth
Glutamine
Leucine
Salmonella
Bacteriophages
Serine
Tyrosine
Amino Acid Sequence
Phenotype
Mutation
endorhamnosidase

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Intragenic suppressors of folding defects in the P22 tailspike protein. / Fane, Bentley A; King, J.

In: Genetics, Vol. 127, No. 2, 1991, p. 263-277.

Research output: Contribution to journalArticle

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