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Science 23 July 1999:
Vol. 285. no. 5427, pp. 591 - 595
DOI: 10.1126/science.285.5427.591
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Reports

Genetic Selection of Peptide Inhibitors of Biological Pathways

Thea C. Norman, 1* Dana L. Smith, 1 Peter K. Sorger, 1 † Becky L. Drees, 2 Sean M. O'Rourke, 3 Timothy R. Hughes, 4 Christopher J. Roberts, 4 Stephen H. Friend, 4 Stan Fields, 2 Andrew W. Murray 1

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.

1 Department of Physiology, University of California, San Francisco, CA 94143-0444, USA.
2 Department of Genetics and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
3 Department of Biochemistry, University of California, San Francisco, CA 94143-0448, USA.
4 Rosetta Inpharmatics, Kirkland, WA 98034, USA.
*   To whom correspondence should be addressed. Present address: Microbia Inc., 840 Memorial Drive, Cambridge, MA 02139, USA. E-mail: tnorman{at}microbia.com

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