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Vol. 9, Issue 1, 89-101, January 1998
Department of Plant Pathology, Cornell University, Ithaca, New York
14853
A gene (NhKIN1) encoding a kinesin was cloned from
Nectria haematococca genomic DNA by polymerase chain
reaction amplification, using primers corresponding to conserved
regions of known kinesin-encoding genes. Sequence analysis showed that
NhKIN1 belongs to the subfamily of conventional kinesins
and is distinct from any of the currently designated kinesin-related
protein subfamilies. Deletion of NhKIN1 by
transformation-mediated homologous recombination caused several dramatic phenotypes: a 50% reduction in colony growth rate, helical or
wavy hyphae with reduced diameter, and subcellular abnormalities including withdrawal of mitochondria from the growing hyphal apex and
reduction in the size of the Spitzenkörper, an apical aggregate of secretory vesicles. The effects on mitochondria and
Spitzenkörper were not due to altered microtubule distribution,
as microtubules were abundant throughout the length of hyphal tip cells
of the mutant. The rate of spindle elongation during anaphase B of
mitosis was reduced 11%, but the rate was not significantly different from that of wild type. This lack of a substantial mitotic phenotype is
consistent with the primary role of the conventional kinesins in
organelle motility rather than mitosis. Our results provide further
evidence that the microtubule-based motility mechanism has a direct
role in apical transport of secretory vesicles and the first evidence
for its role in apical transport of mitochondria in a filamentous
fungus. They also include a unique demonstration that a
microtubule-based motor protein is essential for normal positioning of
the Spitzenkörper, thus providing a new insight into the cellular
basis for the aberrant hyphal morphology.
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