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(Received for publication, September 28,
1995; and in revised form, December 5, 1995) CCA1 codes for mitochondrial, cytosolic, and nuclear
ATP(CTP):tRNA nucleotidyltransferase. Studies reported here examine the
mechanisms leading to and the consequences of altering the distribution
of this important tRNA processing enzyme. We show that the majority of
Cca1p-I, translated from the first in-frame ATG, is in mitochondria but
surprisingly, there is a small contribution to nuclear and cytosolic
tRNA processing by this form as well. The majority of Cca1p-II and
Cca1p-III, translated from ATG2 and ATG3, respectively, is in the
cytosol but both are also located in the nucleus for processing
precursors. Altering the cytosolic/nuclear distribution of Cca1p by
fusing the SV40 nuclear localization signal to the 5` end of CCA1 causes a growth defect and results in the accumulation of
end-shortened tRNAs in the cytosol. These results suggest an important
role for Cca1p in the cytosol of eukaryotes, presumably in the repair
of 3` CCA termini. These experiments also demonstrate that individual
tRNAs are affected differently by reduced cytosolic
nucleotidyltransferase and that cells resuming exponential growth are
more severely affected than those continuing exponential growth.
Volume 271,
Number 9,
Issue of March 1, 1996 pp. 4679-4686
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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