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Vol. 96, Issue 15, 8567-8572, July 20, 1999 (genetic analysis / protein interactions / combinatorial
peptide libraries)
* The Molecular Sciences Institute, 2168 Shattuck Avenue, Berkeley,
CA 94704
Communicated by Sydney Brenner, The Molecular Sciences Institute,
Berkeley, CA, May 20, 1999 (received for review April 6, 1999)
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.
Copyright © 1999 by The National Academy of Sciences 0027-8424/99/968567-6$2.00/0
Genetics
"Mutagenesis" by peptide aptamers identifies genetic
network members and pathway connections
, and
To whom reprint requests should be addressed. e-mail:
colman-lerner{at}molsci.org.
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