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J Biol Chem, Vol. 275, Issue 1, 429-438, January 7, 2000
,
From the Department of Microbiology and Molecular Genetics, Jonsson
Comprehensive Cancer Center, University of California,
Los Angeles, California 90095-1489
Protein farnesyltransferase (FTase) plays
important roles in the growth and differentiation of eukaryotic cells.
In this paper, we report the identification of the
Schizosaccharomyces pombe gene
cpp1+ encoding the 
-subunit of FTase. The
predicted amino acid sequence of the cpp1+ gene
product shares significant similarity with FTase -subunits from a
variety of organisms. S. pombe FTase purified from E. coli exhibits high enzymatic activity toward the CAAX
farnesylation motif substrates (where C represents cysteine,
A represents aliphatic amino acid, and X is
preferentially methionine, cysteine, serine, alanine, or glutamine)
while showing little preference for CAAL geranylgeranylation motif substrates (where L represents
leucine or phenylalanine). cpp1+ is not
essential for growth as shown by gene disruption; however, mutant cells
exhibit rounded or irregular cell morphology. Expression of a
geranylgeranylated mutant form, Ras1-CVIL, which can bypass farnesylation, rescues these morphological defects. We also identify a
novel phenotype of cpp1
mutants,
hypersensitivity to canavanine. This appears to be due to a 3-4-fold
increase in the rate of arginine uptake as compared with wild-type
cells. Expression of the geranylgeranylated mutant form of a novel
farnesylated small GTPase, SpRheb, is able to suppress the elevated
arginine uptake rate. These results demonstrate that protein
farnesylation is critical for maintaining normal cell morphology
through Ras1 and canavanine resistance through SpRheb.
Supported by an Edwin D. Pauley Foundation Fellowship.
§
Supported by United States Public Health Service National Research
Service Award GM07185.
¶
To whom correspondence should be addressed: Dept. of
Microbiology and Molecular Genetics, UCLA, 1602 Molecular Sciences
Bldg., 609 Charles E. Young Dr., Los Angeles, CA 90095-1489. Tel.:
310-206-7318; Fax: 310-206-5231; E-mail:
fuyut@microbio.ucla.edu.
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