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A plant-specific subclass of C-terminal kinesins contains a conserved A-type cyclin-dependent kinase site implicated in folding and dimerization

(2004) PLANT PHYSIOLOGY. 135(3). p.1417-1429
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Abstract
Cyclin-dependent kinases (CDKs) control cell cycle progression through timely coordinated phosphorylation events. Two kmesin-like proteins that interact with CDKA;1 were identified and designated KCA1 and KCA2. They are 81% identical and have a similar three-partite domain organization. The N-terminal domain contains an ATP and microtubule-binding site typical for kinesin motors. A green fluorescent protein (GFP) fusion of the N-terminal domain of KCA1 decorated microtubules in Bright Yellow-2 cells, demonstrating microtubule-binding activity. During cytokinesis the full-length GFP-fusion protein accumulated at the midline of young and mature expanding phragmoplasts. Two-hybrid analysis and coimmunoprecipitation experiments showed that coiled-coil structures of the central stalk were responsible for homo- and heterodimerization of KCA1 and KCA2. By western-blot analysis, high molecular mass KCA molecules were detected in extracts from Bright Yellow-2 cells overproducing the full-length GFP fusion. Treatment of these cultures with the phosphatase inhibitor vanadate caused an accumulation of these KCA molecules. In addition to dimerization, interactions within the C-terminally located tail domain were revealed, indicating that the tail could fold onto itself. The tail domains of KCA1 and KCA2 contained two adjacent putative CDKA;1 phosphorylation sites, one of which is conserved in KCA homologs from other plant species. Site-directed mutagenesis of the conserved phosphorylation sites in KCA1 resulted in a reduced binding with CDKA;1 and abolished intramolecular tail interactions. The data show that phosphorylation of the CDKA;1 site provokes a conformational change in the structure of KCA with implications in folding and dimerization.
Keywords
CELL-CYCLE, CALMODULIN-BINDING PROTEIN, MICROTUBULE DYNAMICS, SPINDLE FORMATION, IN-VIVO, ARABIDOPSIS, MOTOR, PHOSPHORYLATION, CYTOSKELETON, CYTOKINESIS

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Citation

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Chicago
Vanstraelen, Marleen, Juan Antonio Torres Acosta, Lieven De Veylder, Dirk Inzé, and Danny Geelen. 2004. “A Plant-specific Subclass of C-terminal Kinesins Contains a Conserved A-type Cyclin-dependent Kinase Site Implicated in Folding and Dimerization.” Plant Physiology 135 (3): 1417–1429.
APA
Vanstraelen, M., Torres Acosta, J. A., De Veylder, L., Inzé, D., & Geelen, D. (2004). A plant-specific subclass of C-terminal kinesins contains a conserved A-type cyclin-dependent kinase site implicated in folding and dimerization. PLANT PHYSIOLOGY, 135(3), 1417–1429.
Vancouver
1.
Vanstraelen M, Torres Acosta JA, De Veylder L, Inzé D, Geelen D. A plant-specific subclass of C-terminal kinesins contains a conserved A-type cyclin-dependent kinase site implicated in folding and dimerization. PLANT PHYSIOLOGY. 2004;135(3):1417–29.
MLA
Vanstraelen, Marleen, Juan Antonio Torres Acosta, Lieven De Veylder, et al. “A Plant-specific Subclass of C-terminal Kinesins Contains a Conserved A-type Cyclin-dependent Kinase Site Implicated in Folding and Dimerization.” PLANT PHYSIOLOGY 135.3 (2004): 1417–1429. Print.
@article{299671,
  abstract     = {Cyclin-dependent kinases (CDKs) control cell cycle progression through timely coordinated phosphorylation events. Two kmesin-like proteins that interact with CDKA;1 were identified and designated KCA1 and KCA2. They are 81\% identical and have a similar three-partite domain organization. The N-terminal domain contains an ATP and microtubule-binding site typical for kinesin motors. A green fluorescent protein (GFP) fusion of the N-terminal domain of KCA1 decorated microtubules in Bright Yellow-2 cells, demonstrating microtubule-binding activity. During cytokinesis the full-length GFP-fusion protein accumulated at the midline of young and mature expanding phragmoplasts. Two-hybrid analysis and coimmunoprecipitation experiments showed that coiled-coil structures of the central stalk were responsible for homo- and heterodimerization of KCA1 and KCA2. By western-blot analysis, high molecular mass KCA molecules were detected in extracts from Bright Yellow-2 cells overproducing the full-length GFP fusion. Treatment of these cultures with the phosphatase inhibitor vanadate caused an accumulation of these KCA molecules. In addition to dimerization, interactions within the C-terminally located tail domain were revealed, indicating that the tail could fold onto itself. The tail domains of KCA1 and KCA2 contained two adjacent putative CDKA;1 phosphorylation sites, one of which is conserved in KCA homologs from other plant species. Site-directed mutagenesis of the conserved phosphorylation sites in KCA1 resulted in a reduced binding with CDKA;1 and abolished intramolecular tail interactions. The data show that phosphorylation of the CDKA;1 site provokes a conformational change in the structure of KCA with implications in folding and dimerization.},
  author       = {Vanstraelen, Marleen and Torres Acosta, Juan Antonio and De Veylder, Lieven and Inz{\'e}, Dirk and Geelen, Danny},
  issn         = {0032-0889},
  journal      = {PLANT PHYSIOLOGY},
  keyword      = {CELL-CYCLE,CALMODULIN-BINDING PROTEIN,MICROTUBULE DYNAMICS,SPINDLE FORMATION,IN-VIVO,ARABIDOPSIS,MOTOR,PHOSPHORYLATION,CYTOSKELETON,CYTOKINESIS},
  language     = {eng},
  number       = {3},
  pages        = {1417--1429},
  title        = {A plant-specific subclass of C-terminal kinesins contains a conserved A-type cyclin-dependent kinase site implicated in folding and dimerization},
  url          = {http://dx.doi.org/10.1104/pp.104.044818},
  volume       = {135},
  year         = {2004},
}

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