Project: Research project

Project Details


The mitotic spindle is a bipolar protein machine that uses microtubule
(MT) based motors to coordinate chromosome segregation. We propose to
evaluate the function of three MT-associated motor proteins,
KRP130/KLP61F, ncd, and cytoplasmic dynein, with putative mitotic roles
in Drosophila. Based on the ability of these motors to crosslink
microtubules, we hypothesize that they function by mediating
interactions between specifically oriented MTs. Moreover, we
hypothesize that these motors have distinct but possibly overlapping
functions which are likely to be reflected in spatial and temporal
differences in their localization within the mitotic spindle. To test
these hypotheses, we will pursue three specific aims. In the first, we
will use high resolution immunomicroscopy in concert with detailed MT
polarity analyses of Drosophila mitotic spindles to determine the
localization of these motors during different stages of mitosis and the
polarity orientation of the MTs with which they interact. In the
second, we will investigate the MT-MT transport and bundling properties
of the motors in vitro by combining purified motor holoenzymes with
preparations of purified MTs. Particular attention will be given to the
polarity relationships between crosslinked MTs. In the third, we will
study mitotic spindle formation in cell-free extracts immunodepleted of
various mitotic motors. We will then add back purified KRP130/KLP61F,
ncd, and cytoplasmic dynein, individually or in combinations, to
determine their ability to complement the resulting defects in spindle
formation. Collectively, these studies will allow us to assign specific
functions to individual motors and to determine how the functions of
multiple motors are coordinated.
Effective start/end date1/1/9812/31/00


  • Cell Biology


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