BOKU - Universität für Bodenkultur Wien - Forschungsinformationssystem
Gewählte Publikation:
Vanstraelen, M; Torres Acosta, JA; 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 Physiol. 2004; 135(3):1417-1429
FullText
FullText_BOKU
- 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.
- Autor*innen der BOKU Wien:
- Find related publications in this database (using NML MeSH Indexing)
- Amino Acid Sequence -
- Base Sequence -
- Cell Line -
- Cloning, Molecular -
- Conserved Sequence -
- Cyclin-Dependent Kinases - chemistry
- DNA Primers -
- Dimerization -
- Genetic Vectors -
- Kinesin - chemistry
- Molecular Sequence Data -
- Mutagenesis, Site-Directed -
- Peptide Fragments - chemistry
- Plants - cytology
- Polymerase Chain Reaction -
- Protein Biosynthesis - genetics
- Protein Folding -
- Protein Structure, Secondary -
- Recombinant Proteins - chemistry
- Reverse Transcriptase Polymerase Chain Reaction -
- Transcription, Genetic - genetics
Altmetric: