A conserved interaction between Moe1 and Mal3 is important for proper spindle formation in Schizosaccharomyces pombe.
Moe1 is a conserved fission yeast protein that negatively affects microtubule stability/assembly. We conducted a two-hybrid screen to search for Moe1-binding proteins and isolated Mal3, a homologue of human EB1. We show that Moe1 and Mal3 expressed in bacteria form a complex and that Moe1 and Mal3 expressed in fission yeast cosediment with microtubules. Deletion of either moe1 or mal3 does not result in lethality; however, deletion of both moe1 and mal3 leads to cell death in the cold. The resulting cells appear to die of chromosome missegregation, which correlates with the presence of abnormal spindles. We investigated the cause for the formation of monopolar spindles and found that only one of the two spindle pole bodies (SPBs) contains gamma-tubulin, although both SPBs appear to be equal in size and properly inserted in the nuclear membrane. Moreover, the moe1 mal3 double null mutant in the cold contains abnormally short and abundant interphase microtubule bundles. These data suggest that Moe1 and Mal3 play a role in maintaining proper microtubule dynamics/integrity and distribution of gamma-tubulin to the SPBs during mitosis. Finally, we show that human Moe1 and EB1 can each rescue the phenotype of the moe1 mal3 double null mutant and form a complex, suggesting that these proteins are part of a well-conserved mechanism for regulating spindle functioning.
Chen
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Department of Biology, New York University, New York, New York 10003-6688, USA.
Chen
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J
Chang
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D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002467:Cell Nucleus / Q000648:ultrastructure; D039642:Eukaryotic Initiation Factors; D017353:Gene Deletion; D006801:Humans; D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008870:Microtubules / Q000378:metabolism / Q000502:physiology; D012568:Schizosaccharomyces / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D029702:Schizosaccharomyces pombe Proteins; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D014404:Tubulin / Q000378:metabolism
D002352:Carrier Proteins; C405804:EB1 microtubule binding proteins; D039642:Eukaryotic Initiation Factors; D046911:Macromolecular Substances; C108851:Mal3 protein, S pombe; D008869:Microtubule-Associated Proteins; C117671:Moe1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D014404:Tubulin