Abstract
Microtubule plus ends are dynamic ends that interact with other cellular structures. Microtubule plus end tracking proteins are considered to play important roles in the regulation of microtubule plus ends. Recent studies revealed that EB1 is the central regulator for microtubule plus end tracking proteins by recruiting them to microtubule plus ends through direct interaction. Here we report the identification of a novel Drosophila protein, which we call Kebab (kinetochore and EB1 associated basic protein), through in vitro expression screening for EB1-interacting proteins. Kebab fused to GFP shows a novel pattern of dynamic localisation in mitosis. It localises to kinetochores weakly in metaphase and accumulates progressively during anaphase. In telophase, it associates with microtubules in central-spindle and centrosomal regions. The localisation to kinetochores depends on microtubules. The protein has a domain most similar to the atypical CH domain of Ndc80, and a coiled-coil domain. The interaction with EB1 is mediated by two SxIP motifs but is not required for the localisation. Depletion of Kebab in cultured cells by RNA interference did not show obvious defects in mitotic progression or microtubule organisation. Generation of mutants lacking the kebab gene indicated that Kebab is dispensable for viability and fertility.
Highlights
The microtubule cytoskeleton is a dynamic network, constantly reorganising itself in response to various internal and external cues
To gain further insight into the regulation of microtubule plus ends, we have identified a novel EB1-interacting protein, CG31672, that associates with mitotic kinetochores
In this study we identified a novel EB1interating protein, Kebab, which shows a dynamic localisation to kinetochores and microtubules in mitosis
Summary
The microtubule cytoskeleton is a dynamic network, constantly reorganising itself in response to various internal and external cues. In order to perform cellular functions as diverse as chromosome segregation, flagellar movement or neuronal transport, the microtubule network needs complex regulatory mechanisms [1]. Most non-motor MAPs do not have recognisable features within their primary sequence or high sequence conservation across eukaryotes. As hundreds of MAPs interact with microtubules even in a single cell, functional redundancies are likely to be very high. As the behaviour of microtubules varies within cells, and in different cell cycle stages and cell types, MAPs must be spatially and temporally regulated. We are still a long way from knowing the full complement of MAPs, how they regulate microtubules, and how they themselves are regulated in cells
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