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J. Biol. Chem., Vol. 276, Issue 9, 6392-6397, March 2, 2001
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From the Department of Molecular Biology and Genetics, Cornell
University, Ithaca, New York 14853-2703
The protein specified by the Saccharomyces
cerevisiae nuclear gene PET111 specifically activates
translation of the mitochondrially coded mRNA for cytochrome
c oxidase subunit II (Cox2p). We found Pet111p specifically
in mitochondria of both wild-type cells and cells expressing a
chromosomal gene for a functional epitope-tagged form of Pet111p.
Pet111p was associated with mitochondrial membranes and was highly
resistant to extraction with alkaline carbonate. Pet111p was protected
from proteolytic digestion by the mitochondrial inner membrane. Thus,
it is exposed only on the matrix side, where it could participate
directly in organellar translation and localize Cox2p synthesis by
virtue of its functional interaction with the COX2 mRNA
5'-untranslated leader. We also found that Pet111p is present at levels
limiting the synthesis of Cox2p by examining the effect of altered
PET111 gene dosage in the nucleus on expression of a
reporter gene, cox2::ARG8m, that was
inserted into mitochondrial DNA. The level of the reporter protein,
Arg8p, was one-half that of wild type in a diploid strain heterozygous
for a pet111 deletion mutation, whereas it was increased 2.8-fold in a strain bearing extra copies of PET111 on a
high-copy plasmid. Thus, Pet111p could play dual roles in both membrane localization and regulation of Cox2p synthesis within mitochondria.
Pet111p, an Inner Membrane-bound Translational Activator That
Limits Expression of the Saccharomyces cerevisiae
Mitochondrial Gene COX2*
, and
*
This investigation was supported by National Institutes of
Health Research Grant GM29362 and Training Grant GM07617.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Present address: Invitrogen Corp., 1600 Faraday Ave., Carlsbad, CA 92008.
§
To whom correspondence should be addressed: Dept. of Molecular
Biology and Genetics, Biotechnology Bldg., Cornell University, Ithaca,
New York 14853-2703. Tel.: 607-254-4835; Fax: 607-255-6249; E-mail:
tdf1@cornell.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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