Genetic selection of peptide inhibitors of biological pathways.
Genetic selections were used to find peptides that inhibit biological pathways in budding yeast. The peptides were presented inside cells as peptamers, surface loops on a highly expressed and biologically inert carrier protein, a catalytically inactive derivative of staphylococcal nuclease. Peptamers that inhibited the pheromone signaling pathway, transcriptional silencing, and the spindle checkpoint were isolated. Putative targets for the inhibitors were identified by a combination of two-hybrid analysis and genetic dissection of the target pathways. This analysis identified Ydr517w as a component of the spindle checkpoint and reinforced earlier indications that Ste50 has both positive and negative roles in pheromone signaling. Analysis of transcript arrays showed that the peptamers were highly specific in their effects, which suggests that they may be useful reagents in organisms that lack sophisticated genetics as well as for identifying components of existing biological pathways that are potential targets for drug discovery.
Norman
T C
TC
Department of Physiology, University of California, San Francisco, CA 94143-0444, USA. tnorman@microbia.com
Smith
D L
DL
Sorger
P K
PK
Drees
B L
BL
O'Rourke
S M
SM
Hughes
T R
TR
Roberts
C J
CJ
Friend
S H
SH
Fields
S
S
Murray
A W
AW
D000595:Amino Acid Sequence; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D016193:G1 Phase; D005690:Galactose / Q000378:metabolism; D008074:Lipoproteins / Q000378:metabolism; D000072235:Mating Factor; D008836:Micrococcal Nuclease; D008938:Mitosis; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D010675:Pheromones / Q000378:metabolism*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D012641:Selection, Genetic; D015398:Signal Transduction; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism*; D014158:Transcription, Genetic
D005656:Fungal Proteins; D008074:Lipoproteins; C095063:MFA2 protein, S cerevisiae; D019151:Peptide Library; D010455:Peptides; D010675:Pheromones; C079506:STE50 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D000072235:Mating Factor; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; C093416:MPS1 protein, S cerevisiae; D008836:Micrococcal Nuclease; D005690:Galactose