Drosophila Nod Protein Binds Preferentially to the Plus Ends of Microtubules and Promotes Microtubule Polymerization In Vitro super

Nod, a nonmotile kinesinlike protein, plays a critical role in segregating achiasmate chromosomes during female meiosis. In addition to localizing to oocyte chromosomes, we show that functional full-length Nod-GFP (Nod sub(FL)-GFP) localizes to the posterior pole of the oocyte at stages 9-10A, as do...

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Bibliographic Details
Published in:Molecular biology of the cell 2005-11, Vol.16 (11), p.5400-5409.
Main Authors: Cui, Wei, Sproul, Lisa R, Gustafson, Susan M, Matthies, Heinrich JG, Gilbert, Susan P, Hawley, R S
Format: Article
Language:English
Subjects:
Drosophila
Escherichia coli
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Summary:Nod, a nonmotile kinesinlike protein, plays a critical role in segregating achiasmate chromosomes during female meiosis. In addition to localizing to oocyte chromosomes, we show that functional full-length Nod-GFP (Nod sub(FL)-GFP) localizes to the posterior pole of the oocyte at stages 9-10A, as does kinesin heavy chain (KHC), a plus end-directed motor. This posterior localization is abolished in grk mutants that no longer maintain the microtubule (MT) gradient in the oocyte. To test the hypothesis that Nod binds to the plus ends of MTs, we expressed and purified both full-length Nod (Nod sub(FL)) and a truncated form of Nod containing only the motorlike domain (Nod sub(318)) from Escherichia coli and assessed their interactions with MTs in vitro. Both Nod sub(FL) and Nod sub(318) demonstrate preferential binding to the ends of the MTs, displaying a strong preference for binding to the plus ends. When Nod sub(318)-GFP:MT collision complexes were trapped by glutaraldehyde fixation, the preference for binding to plus ends versus minus ends was 17:1. Nod sub(FL) and Nod sub(318) also promote MT polymerization in vitro in a time-dependent manner. The observation that Nod is preferentially localized to the plus ends of MTs and stimulates MT polymerization suggests a mechanism for its function. hr> Articles from Molecular Biology of the Cell are provided here courtesy of American Society for Cell Biology Write to PMC A |A PMC Home A |A PubMed NCBI A |A U.S. National Library of Medicine NIH A |A Department of Health and Human Services Privacy Policy A |A Disclaimer A |A Freedom of Information Act
ISSN:1059-1524
1059-1524
DOI:10.1091/mbc.E05-06-0582