"Mutagenesis" by peptide aptamers identifies genetic network members and pathway connections

Proc Natl Acad Sci U S A. 1999 Jul 20;96(15):8567-72. doi: 10.1073/pnas.96.15.8567.

Abstract

We selected peptide aptamers from combinatorial libraries that disrupted cell-cycle arrest caused by mating pheromone in yeast. We used these aptamers as baits in two-hybrid hunts to identify genes involved in cell-cycle arrest. These experiments identified genes known to function in the pathway, as well as a protein kinase, the CBK1 product, whose function was not known. We used a modified two-hybrid system to identify specific interactions disrupted by these aptamers. These experiments demonstrate a means to perform "genetics" on the protein complement of a cell without altering its genetic material. Peptide aptamers can be identified that disrupt a process. These aptamers can then be used as affinity reagents to identify individual proteins and protein interactions needed for the process. Forward genetic analysis with peptide aptamer "mutagens" should be particularly useful in elucidating genetic networks in organisms and processes for which classical genetics is not feasible.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Cell Cycle / drug effects*
  • Cell Cycle / genetics
  • Cell Division / drug effects
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Genes, Fungal
  • Mating Factor
  • Molecular Sequence Data
  • Mutagenesis*
  • Peptide Library
  • Peptides / chemistry
  • Peptides / pharmacology*
  • Saccharomyces cerevisiae / genetics*
  • Sequence Deletion

Substances

  • Fungal Proteins
  • Peptide Library
  • Peptides
  • Mating Factor