In-frame linker insertion mutagenesis of yeast transposon Ty1: phenotypic analysis

L. T. Braiterman, G. M. Monokian, D. J. Eichinger, S. L. Merbs, A. Gabriel, J. D. Boeke

Research output: Contribution to journalArticlepeer-review

Abstract

A plasmid bearing a transpositionally functional GAL1::Tyl fusion was mutagenized by insertion of four or five codons semirandomly throughout the plasmid. This collection of mutant plasmids was introduced into yeast cells and studied with regard to the properties of the mutant Ty1-encoded proteins and the transposition phenotypes observed. All of the transposition-inactivating mutations were previously found to be recessive with the exception of a single mutation in TYA. In this mutant, TYA protein of normal abundance is produced, but the virus-like particles containing this protein are unstable and have aberrant behavior. The effects of mutations in noncoding regions, as well as the capsid protein coding region TYA, and the regions encoding the protease, integrase and reverse transcriptase proteins are described. Effects on gene expression, types of proteins produced, proteolysis of precursor proteins, virus-like particle structure, and biochemical activities of the encoded proteins are summarized. In addition, we show that one of the mutations in the 3' LTR represents a new nonessential site into which foreign marker DNA can be inserted without compromising transposition.

Original languageEnglish (US)
Pages (from-to)19-26
Number of pages8
JournalGene
Volume139
Issue number1
DOIs
StatePublished - Feb 11 1994

Keywords

  • Retrotransposons
  • Saccharomyces cerevisiae
  • codon insertion mutations
  • integrase
  • protease
  • reverse transcriptase

ASJC Scopus subject areas

  • Genetics

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