Arabidopsis SPIRAL2 promotes uninterrupted microtubule growth by suppressing the pause state of microtubule dynamics.
SPIRAL2 (SPR2) of Arabidopsis thaliana is a microtubule-associated protein containing multiple HEAT repeats that are found only in the plant lineage. We show that SPR2 and SP2L, their closest Arabidopsis homolog, are expressed in various tissues with partially overlapping patterns, and spr2-sp2l double mutants exhibit enhanced right-handed helical growth. Fusion to green fluorescent protein (GFP) expressed under the control of the native regulatory elements showed that both SPR2 and SP2L were localized to cortical microtubules, mainly in particles of various sizes. Along the microtubule, the GFP-fused forms also distributed partly at the plus ends. In the spr2-mutant background, cortical microtubules were less dynamic, and the pause state - in which microtubules undergo neither growth nor shrinkage - increased at the plus ends. The continuous plus-end tracking of GFP-EB1 was occasionally interrupted in the mutant cells. Recombinant SPR2 protein promoted microtubule polymerization, and bound to microtubules with an N-terminal segment that contained two HEAT repeats as well as to those with a C-terminal region. In vitro analyses of microtubule dynamics revealed that SPR2 and SP2L suppressed the pause state at microtubule ends, thereby leading to enhanced microtubule growth. We propose that the SPR2-family proteins act on the pause state to facilitate a transition to microtubule growth.
Yao
Maki
M
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Nara, Japan.
Wakamatsu
Yoshinori
Y
Itoh
Tomohiko J
TJ
Shoji
Tsubasa
T
Hashimoto
Takashi
T
D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000254:growth & development*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018546:Hypocotyl / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D036226:Seedlings / Q000378:metabolism
D029681:Arabidopsis Proteins; C483461:EB1 protein, Arabidopsis; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; C496076:SPIRAL2 protein, Arabidopsis; D049452:Green Fluorescent Proteins