QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Vol. 16, Issue 11, 5400-5409, November 2005

This Article Full Text Full Text (PDF) Supplemental Figure All Versions of this Article: E05-06-0582v1 16/11/5400    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Cui, W. Articles by Hawley, R. S. Search for Related Content PubMed PubMed Citation Articles by Cui, W. Articles by Hawley, R. S.

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

Wei Cui ^*  , Lisa R. Sproul  , Susan M. Gustafson ^*, Heinrich J.G. Matthies ^||, Susan P. Gilbert , and R. S. Hawley ^* ¶

^* Stowers Institute for Medical Research, Kansas City, MO 64110; Department of Pathology and Clinical Laboratory, University of Kansas Medical Center, Kansas City, KS 66160; Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260; ^|| Department of Biological Sciences, Vanderbilt University, Nashville, TN 37235; and ^¶ Department of Molecular Biosciences, University of Kansas, Lawrence, KS 66045

Submitted June 29, 2005; Revised August 12, 2005; Accepted August 25, 2005
Monitoring Editor: J. Richard McIntosh

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_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_FL) and a truncated form of Nod containing only the motorlike domain (Nod₃₁₈) from Escherichia coli and assessed their interactions with MTs in vitro. Both Nod_FL and Nod₃₁₈ demonstrate preferential binding to the ends of the MTs, displaying a strong preference for binding to the plus ends. When Nod₃₁₈-GFP:MT collision complexes were trapped by glutaraldehyde fixation, the preference for binding to plus ends versus minus ends was 17:1. Nod_FL and Nod₃₁₈ 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.

Abbreviations used: MT, microtubule; GFP, green fluorescent protein; KHC, kinesin heavy chain; NEB, nuclear envelope breakdown.

The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

These authors contributed equally to this work.

Address correspondence to: R. Scott Hawley (rsh{at}stowers-institute.org).

This article has been cited by other articles:

				Y. Xiang and R. S. Hawley The Mechanism of Secondary Nondisjunction in Drosophila melanogaster Females Genetics, September 1, 2006; 174(1): 67 - 78. [Abstract] [Full Text] [PDF]

				R. S. Shapiro and K. V. Anderson Drosophila Ik2, a member of the I{kappa}B kinase family, is required for mRNA localization during oogenesis Development, April 15, 2006; 133(8): 1467 - 1475. [Abstract] [Full Text] [PDF]

				W. Cui and R. S. Hawley The HhH(2)/NDD Domain of the Drosophila Nod Chromokinesin-like Protein Is Required for Binding to Chromosomes in the Oocyte Nucleus Genetics, December 1, 2005; 171(4): 1823 - 1835. [Abstract] [Full Text] [PDF]