Nuclear-localized subtype of end-binding 1 protein regulates spindle organization in Arabidopsis.
End-binding 1 (EB1) proteins are evolutionarily conserved plus-end-tracking proteins that localize to growing microtubule plus ends where they regulate microtubule dynamics and interactions with intracellular targets. Animal EB1 proteins have acidic C-terminal tails that might induce an autoinhibitory conformation. Although EB1 proteins with the same structural features occur in plants (EB1a and EB1b in Arabidopsis thaliana), a variant form (EB1c) is present that lacks the characteristic tail. We show that in Arabidopsis the tail region of EB1b, but not of EB1c, inhibits microtubule assembly in vitro. EB1a and EB1b form heterodimers with each other, but not with EB1c. Furthermore, the EB1 genes are expressed in various cell types of Arabidopsis, but the expression of EB1c is particularly strong in the meristematic cells where it is targeted to the nucleus by a nuclear localization signal in the C-terminal tail. Reduced expression of EB1c compromised the alignment of spindle and phragmoplast microtubules and caused frequent lagging of separating chromosomes at anaphase. Roots of the eb1c mutant were hypersensitive to a microtubule-disrupting drug and complete rescue of the mutant phenotype required the tail region of EB1c. These results suggest that a plant-specific EB1 subtype has evolved to function preferentially on the spindle microtubules by accumulating in the prophase nucleus.
Komaki
Shinichiro
S
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630-0192, Japan.
Abe
Tatsuya
T
Coutuer
Silvie
S
Inzé
Dirk
D
Russinova
Eugenia
E
Hashimoto
Takashi
T
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D020090:Chromosome Segregation / Q000235:genetics / Q000502:physiology; D004136:Dinitrobenzenes / Q000494:pharmacology; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D010641:Phenotype; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D008941:Spindle Apparatus / Q000378:metabolism*; D013424:Sulfanilamides / Q000494:pharmacology; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics / Q000378:metabolism
D029681:Arabidopsis Proteins; D004136:Dinitrobenzenes; C483461:EB1 protein, Arabidopsis; D013424:Sulfanilamides; C012465:oryzalin