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Molecular and Cellular Biology, October 2001, p. 7035-7046, Vol. 21, No. 20
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.20.7035-7046.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Induction of Distinct [URE3] Yeast
Prion Strains
Martin
Schlumpberger,1,2
Stanley B.
Prusiner,1,2,3,* and
Ira
Herskowitz3
Institute for Neurodegenerative
Diseases1 and Departments of
Neurology2 and Biochemistry and
Biophysics,3 University of California, San
Francisco, California 94143-0518
Received 7 May 2001/Returned for modification 4 June 2001/Accepted 18 July 2001
[URE3] is a non-Mendelian genetic element in
Saccharomyces cerevisiae, which is caused by a
prion-like, autocatalytic conversion of the Ure2 protein (Ure2p) into
an inactive form. The presence of [URE3] allows yeast
cells to take up ureidosuccinic acid in the presence of ammonia. This
phenotype can be used to select for the prion state. We have developed
a novel reporter, in which the ADE2 gene is controlled
by the DAL5 regulatory region, which allows monitoring
of Ure2p function by a colony color phenotype. Using this reporter, we
observed induction of different [URE3] prion variants
("strains") following overexpression of the N-terminal Ure2p
prion domain (UPD) or full-length Ure2p. Full-length Ure2p induced two
types of [URE3]: type A corresponds to conventional [URE3], whereas the novel type B variant is
characterized by relatively high residual Ure2p activity and efficient
curing by coexpression of low amounts of a UPD-green fluorescent
protein fusion protein. Overexpression of UPD induced type B
[URE3] but not type A. Both type A and B
[URE3] strains, as well as weak and strong isolates of
type A, were shown to stably maintain different prion strain characteristics. We suggest that these strain variants result from
different modes of aggregation of similar Ure2p monomers. We also
demonstrate a procedure to counterselect against the
[URE3] state.
*
Corresponding author. Mailing address: Institute for
Neurodegenerative Diseases, Box 0518, University of California, San
Francisco, CA 94143-0518. Phone: (415) 476-4482. Fax: (415) 476-8386. E-mail: stanley{at}itsa.ucsf.edu.
Molecular and Cellular Biology, October 2001, p. 7035-7046, Vol. 21, No. 20
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.20.7035-7046.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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