"Mutagenesis" by peptide aptamers identifies genetic network members and pathway connections.
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.
The Molecular Sciences Institute, 2168 Shattuck Avenue, Berkeley, CA 94704, USA.
C R
CR
Geyer
A
A
Colman-Lerner
R
R
Brent
D000595:Amino Acid Sequence; D002453:Cell Cycle / Q000187:drug effects* / Q000235:genetics; D002455:Cell Division / Q000187:drug effects; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal; D000072235:Mating Factor; D008969:Molecular Sequence Data; D016296:Mutagenesis; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017384:Sequence Deletion
D005656:Fungal Proteins; D019151:Peptide Library; D010455:Peptides; D000072235:Mating Factor