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Vol. 9, Issue 10, 2839-2855, October 1998
and
Department of Molecular, Cellular, and Developmental Biology,
University of Colorado, Boulder, Colorado 80309-0347
The "cut" mutants of Schizosaccharomyces pombe
are defective in spindle formation and/or chromosome segregation, but
they proceed through the cell cycle, resulting in lethality. Analysis of temperature-sensitive alleles of cut11+
suggests that this gene is required for the formation of a functional bipolar spindle. Defective spindle structure was revealed with fluorescent probes for tubulin and DNA. Three-dimensional
reconstruction of mutant spindles by serial sectioning and electron
microscopy showed that the spindle pole bodies (SPBs) either failed to
complete normal duplication or were free floating in the nucleoplasm.
Localization of Cut11p tagged with the green fluorescent protein showed
punctate nuclear envelope staining throughout the cell cycle and SPBs
staining from early prophase to mid anaphase. This SPB localization
correlates with the time in the cell cycle when SPBs are inserted into
the nuclear envelope. Immunoelectron microscopy confirmed the
localization of Cut11p to mitotic SPBs and nuclear pore
complexes. Cloning and sequencing showed that
cut11+ encodes a novel protein with seven
putative membrane-spanning domains and homology to the
Saccharomyces cerevisiae gene NDC1. These
data suggest that Cut11p associates with nuclear pore complexes and
mitotic SPBs as an anchor in the nuclear envelope; this role is
essential for mitosis.
Present addresses: Children's Hospital, 1050 Enders Building, 300 Longwood Avenue, Boston, MA 02115.
Imperial Cancer Research Fund, 123 Lincoln's Inn Fields, London, United Kingdom.
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