Use of GFP Fusions to a Microtubule Motor Protein to Analyze Spindle Dynamics in Live Oocytes and Embryos of Drosophila

1997 ◽  
Vol 3 (S2) ◽  
pp. 127-128
Author(s):  
S. A. Endow ◽  
D. J. Komma
Keyword(s):  
Fusion Proteins ◽  
Motor Protein ◽  
Spindle Motor ◽  
Gene Fusions ◽  
Mitotic Spindles ◽  
Wild Type ◽  
Early Embryos ◽  
Microtubule Motor ◽  
Microtubule Motor Protein ◽  
Spindle Fibers

Ncd is a kinesin-related microtubule motor protein of Drosophila that plays essential roles in spindle assembly and function during meiosis in oocytes and mitosis in early embryos. Antibody staining experiments have localized the Ned motor protein to spindle fibers and spindle poles throughout the meiotic and early mitotic divisions, demonstrating that Ncd is a spindle motor.We have made ncd-gfp gene fusions with wild-type and S65T gfp and expressed the chimaeric genes in Drosophila to target GFP to the spindle. Transgenic Drosophila carrying the ncd-gfp gene fusions in an ncd null mutant background are wild type with respect to chromosome segregation, indicating that the Ncd-GFP fusion proteins can replace the function of wild-type Ncd. The Ncd-GFP fusion proteins in transgenic Drosophila are expressed under the regulation of the native ncd promoter.Analysis of live Drosophila oocytes and early embryos shows green fluorescent spindles, demonstrating association of Ncd-GFP with meiotic and mitotic spindles. In mitotic spindles, Ncd-GFP localizes to centrosomes (Fig. 1a) and spindle fibers (Fig. 1b).

Mutants of the microtubule motor protein, nonclaret disjunctional, affect spindle structure and chromosome movement in meiosis and mitosis

1992 ◽  
Vol 101 (3) ◽  
pp. 547-559 ◽  
Author(s):  
M. Hatsumi ◽  
S.A. Endow
Keyword(s):  
Motor Protein ◽  
Spindle Motor ◽  
Meiotic Spindle ◽  
Chromosome Movement ◽  
Chromosome Distribution ◽  
Early Embryos ◽  
Microtubule Motor ◽  
Microtubule Motor Protein ◽  
Spindle Structure ◽  
Meiosis I

The Drosophila microtubule motor protein, nonclaret disjunctional (ncd), is required for proper chromosome distribution in meiosis and mitosis. We have examined the meiotic and mitotic divisions in wild-type Drosophila oocytes and early embryos, and the effects of three ncd mutants (cand, ncd and ncdD) on spindle structure and chromosome movement. The ncd mutants cause abnormalities in spindle structure early in meiosis I, and abnormal chromosome configurations throughout meiosis I and II. Defective divisions continue in early embryos of the motor null mutant, cand, with abnormal early mitotic spindles. The effects of mutants on spindle structure suggest that ncd is required for proper meiotic spindle assembly, and may play a role in forming or maintaining spindle poles in meiosis. The disruption of normal meiotic and mitotic chromosome distribution by ncd mutants can be attributed to its role as a spindle motor, although a role for ncd as a chromosome-associated motor protein is not excluded. The ncd motor protein functions not only in meiosis, but also performs an active role in the early mitotic divisions of the embryo.

The Drosophila ncd microtubule motor protein is spindle-associated in meiotic and mitotic cells

1992 ◽  
Vol 103 (4) ◽  
pp. 1013-1020 ◽  
Author(s):  
M. Hatsumi ◽  
S.A. Endow
Keyword(s):  
Sequence Similarity ◽  
Motor Protein ◽  
Spindle Motor ◽  
In Vitro Motility ◽  
Early Embryos ◽  
Microtubule Motor ◽  
Spindle Function ◽  
Microtubule Motor Protein ◽  
Mitotic Cells

The nonclaret disjunctional (ncd) protein is required for normal chromosome distribution in oocytes and early embryos. Mutants of ncd cause frequent nondisjunction and loss of chromosomes, suggesting a role for the protein in spindle function or chromosome movement in meiosis and early mitosis. The ncd protein contains a region of predicted sequence similarity to the microtubule motor protein, kinesin. In vitro motility assays have demonstrated that ncd is a motor that unexpectedly moves toward the minus ends of microtubules, opposite to the direction of kinesin movement. Using antibodies directed against nonconserved regions of the protein, we have localized the ncd motor protein to the meiotic and early mitotic spindle, and to spindles in a mitotically dividing cultured cell line. Its presence in the spindle of meiotic and mitotic cells implies a role for the protein as a spindle motor. The motor may play an essential role in establishing spindle bipolarity in meiosis.

scholarly journals The organization of Golgi in Drosophila bristles requires microtubule motor protein function and a properly organized microtubule array

PLoS ONE ◽  
2019 ◽  
Vol 14 (10) ◽  
pp. e0223174
Author(s):  
Anna Melkov ◽  
Raju Baskar ◽  
Rotem Shachal ◽  
Yehonathan Alcalay ◽  
Uri Abdu
Keyword(s):  
Protein Function ◽  
Motor Protein ◽  
Microtubule Array ◽  
Microtubule Motor ◽  
Microtubule Motor Protein

Organisation and structure of microtubules and microtubule-motor protein complexes

1998 ◽  
Vol 27 (5) ◽  
pp. 446-454 ◽  
Author(s):  
R. H. Wade ◽  
P. Meurer-Grob ◽  
F. Metoz ◽  
I. Arnal
Keyword(s):  
Protein Complexes ◽  
Motor Protein ◽  
Microtubule Motor ◽  
Microtubule Motor Protein

scholarly journals Yeast Kar3 is a minus-end microtubule motor protein that destabilizes microtubules preferentially at the minus ends.

1994 ◽  
Vol 13 (11) ◽  
pp. 2708-2713 ◽  
Author(s):  
S.A. Endow ◽  
S.J. Kang ◽  
L.L. Satterwhite ◽  
M.D. Rose ◽  
V.P. Skeen ◽  
...  
Keyword(s):  
Motor Protein ◽  
Microtubule Motor ◽  
Microtubule Motor Protein

scholarly journals Multiscale modeling and simulation of microtubule–motor-protein assemblies

2015 ◽  
Vol 92 (6) ◽  
Author(s):  
Tong Gao ◽  
Robert Blackwell ◽  
Matthew A. Glaser ◽  
M. D. Betterton ◽  
Michael J. Shelley
Keyword(s):  
Modeling And Simulation ◽  
Multiscale Modeling ◽  
Motor Protein ◽  
Protein Assemblies ◽  
Microtubule Motor ◽  
Microtubule Motor Protein

scholarly journals Chemical subdomains within the kinetochore domain of isolated CHO mitotic chromosomes.

1991 ◽  
Vol 114 (2) ◽  
pp. 285-294 ◽  
Author(s):  
L Wordeman ◽  
E R Steuer ◽  
M P Sheetz ◽  
T Mitchison
Keyword(s):  
Indirect Immunofluorescence ◽  
Motor Protein ◽  
Chromosome Translocation ◽  
Mitotic Chromosomes ◽  
Cell Biol ◽  
Microtubule Motor ◽  
Specific Antigens ◽  
Microtubule Motor Protein

We have used indirect immunofluorescence in combination with correlative EM to subdivide the mammalian kinetochore into two domains based on the localization of specific antigens. We demonstrate here that the fibrous corona on the distal face of the kinetochore plate contains tubulin (previously shown by Mitchison, T. J., and M. W. Kirschner. 1985. J. Cell Biol. 101:755-765) and the minus end-directed, ATP-dependent microtubule motor protein, dynein; whereas a 50-kD CREST antigen is located internal to these components in the kinetochore. Tubulin and dynein can be extracted from the kinetochore by 150 mM KI, leaving other, as yet uncharacterized, components of the kinetochore corona intact. Microtubules and tubulin subunits will associate with kinetochores in vitro after extraction with 150 mM KI, suggesting that other functionally significant, corona-associated molecules remain unextracted. Our results suggest that the corona region of the kinetochore contains the machinery for chromosome translocation along microtubules.

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