Plant γ-tubulin interacts with αβ-tubulin dimers and forms membrane-associated complexes

被引:95
作者
Dryková, D
Cenklová, V
Sulimenko, V
Volc, J
Dráber, P
Binarová, P
机构
[1] Acad Sci Czech Republ, Inst Microbiol, CR-14220 Prague 4, Czech Republic
[2] Acad Sci Czech Republ, Inst Expt Bot, Olomouc 77200, Czech Republic
[3] Acad Sci Czech Republ, Inst Mol Genet, CR-14220 Prague 4, Czech Republic
关键词
D O I
10.1105/tpc.007005
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
gamma-Tubulin is assumed to participate in microtubule nucleation in acentrosomal plant cells, but the underlying molecular mechanisms are still unknown. Here, we show that gamma-tubulin is present in protein complexes of various sizes and different subcellular locations in Arabidopsis and fava bean. Immunoprecipitation experiments revealed an association of gamma-tubulin with alphabeta-tubulin dimers. gamma-Tubulin cosedimented with microtubules polymerized in vitro and localized along their whole length. Large gamma-tubulin complexes resistant to salt treatment were found to be associated with a highspeed microsomal fraction. Blue native electrophoresis of detergent-solubilized microsomes showed that the molecular mass of the complexes was > 1 MD. Large gamma-tubulin complexes were active in microtubule nucleation, but nucleation activity was not observed for the smaller complexes. Punctate gamma-tubulin staining was associated with microtubule arrays, accumulated with short kinetochore microtubules interacting in polar regions with membranes, and localized in the vicinity of nuclei and in the area of cell plate formation. Our results indicate that the association of gamma-tubulin complexes with dynamic membranes might ensure the flexibility of noncentrosomal microtubule nucleation. Moreover, the presence of other molecular forms of gamma-tubulin suggests additional roles for this protein species in microtubule organization.
引用
收藏
页码:465 / 480
页数:16
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