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*Pasteur Institute-Cenci Bolognetti Foundation, Department of Cell
and Developmental Biology, University of Rome "La Sapienza", 00185 Rome, Italy;
We report here the functional characterization of an essential
Saccharomyces cerevisiae gene, MPR1,
coding for a regulatory proteasomal subunit for which the name Rpn11p
has been proposed. For this study we made use of the
mpr1-1 mutation that causes the following pleiotropic
defects. At 24°C growth is delayed on glucose and impaired on
glycerol, whereas no growth is seen at 36°C on either carbon source.
Microscopic observation of cells growing on glucose at 24°C shows
that most of them bear a large bud, whereas mitochondrial morphology is
profoundly altered. A shift to the nonpermissive temperature produces
aberrant elongated cell morphologies, whereas the nucleus fails to
divide. Flow cytometry profiles after the shift to the nonpermissive
temperature indicate overreplication of both nuclear and mitochondrial
DNA. Consistently with the identification of Mpr1p with a proteasomal
subunit, the mutation is complemented by the human POH1
proteasomal gene. Moreover, the mpr1-1 mutant grown to
stationary phase accumulates ubiquitinated proteins. Localization of
the Rpn11p/Mpr1p protein has been studied by green fluorescent
protein fusion, and the fusion protein has been found to be
mainly associated to cytoplasmic structures. For the first time, a
proteasomal mutation has also revealed an associated mitochondrial
phenotype. We actually showed, by the use of [rho°]
cells derived from the mutant, that the increase in DNA content per
cell is due in part to an increase in the amount of mitochondrial DNA.
Moreover, microscopy of mpr1-1 cells grown on glucose
showed that multiple punctate mitochondrial structures were present in
place of the tubular network found in the wild-type strain. These data
strongly suggest that mpr1-1 is a valuable tool with
which to study the possible roles of proteasomal function in
mitochondrial biogenesis.
Department of General Physiology and
Biochemistry, University of Milan, 20133 Milan, Italy; and
§Laboratoire de Génétique Moléculaire
Bâtiment 400, Université Paris-Sud, 91405 Orsay,
France
Corresponding author. E-mail address:
Rinaldit{at}axcasp.caspur.it.
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