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Vol. 12, Issue 1, 221-238, January 2001
Molecular Membrane Biology Laboratory, RIKEN, 2-1 Wako, Saitama
351-0198, Japan
ADP-ribosylation factors, a family of small GTPases, are believed
to be key regulators of intracellular membrane traffic. However, many
biochemical in vitro experiments have led to different models for their
involvement in various steps of vesicular transport, and their precise
role in living cells is still unclear. We have taken advantage of the
powerful yeast genetic system and screened for temperature-sensitive
(ts) mutants of the ARF1 gene from Saccharomyces cerevisiae. By random mutagenesis of the whole open reading
frame of ARF1 by error-prone PCR, we isolated eight
mutants and examined their phenotypes. arf1 ts mutants
showed a variety of transport defects and morphological alterations in
an allele-specific manner. Furthermore, intragenic complementation was
observed between certain pairs of mutant alleles, both for cell growth
and intracellular transport. These results demonstrate that the single
Arf1 protein is indeed involved in many different steps of
intracellular transport in vivo and that its multiple roles may be
dissected by the mutant alleles we constructed.
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