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PMID:14688294

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Citation

Reddy, VS, Day, IS, Thomas, T and Reddy, AS (2004) KIC, a novel Ca2+ binding protein with one EF-hand motif, interacts with a microtubule motor protein and regulates trichome morphogenesis. Plant Cell 16:185-200

Abstract

Kinesin-like calmodulin binding protein (KCBP) is a microtubule motor protein involved in the regulation of cell division and trichome morphogenesis. Genetic studies have shown that KCBP is likely to interact with several other proteins. To identify KCBP-interacting proteins, we used the C-terminal region of KCBP in a yeast two-hybrid screen. This screening resulted in the isolation of a novel KCBP-interacting Ca2+ binding protein (KIC). KIC, with its single EF-hand motif, bound Ca2+ at a physiological concentration. Coprecipitation with bacterially expressed protein and native KCBP, gel-mobility shift studies, and ATPase assays with the KCBP motor confirmed that KIC interacts with KCBP in a Ca2+-dependent manner. Interestingly, although both Ca2+-KIC and Ca2+-calmodulin were able to interact with KCBP and inhibit its microtubule binding activity, the concentration of Ca2+ required to inhibit the microtubule-stimulated ATPase activity of KCBP by KIC was threefold less than that required for calmodulin. Two KIC-related Ca2+ binding proteins and a centrin from Arabidopsis, which contain one and four EF-hand motifs, respectively, bound Ca2+ but did not affect microtubule binding and microtubule-stimulated ATPase activities of KCBP, indicating the specificity of Ca2+ sensors in regulating their targets. Overexpression of KIC in Arabidopsis resulted in trichomes with reduced branch number resembling the zwichel/kcbp phenotype. These results suggest that KIC modulates the activity of KCBP in response to changes in cytosolic Ca2+ and regulates trichome morphogenesis.

Links

PubMed PMC301404 Online version:10.1105/tpc.016600

Keywords

Adenosine Triphosphatases/antagonists & inhibitors; Adenosine Triphosphatases/metabolism; Amino Acid Sequence; Arabidopsis/genetics; Arabidopsis/growth & development; Arabidopsis/metabolism; Arabidopsis Proteins/genetics; Arabidopsis Proteins/isolation & purification; Arabidopsis Proteins/metabolism; Calcium/metabolism; Calcium-Binding Proteins/genetics; Calcium-Binding Proteins/isolation & purification; Calcium-Binding Proteins/metabolism; Calmodulin/metabolism; Calmodulin-Binding Proteins/genetics; Calmodulin-Binding Proteins/metabolism; Cell Surface Extensions/genetics; Cell Surface Extensions/physiology; EF Hand Motifs/genetics; Gene Expression Regulation, Developmental; Gene Expression Regulation, Plant; Microtubule-Associated Proteins/genetics; Microtubule-Associated Proteins/isolation & purification; Microtubule-Associated Proteins/metabolism; Molecular Sequence Data; Phylogeny; Protein Interaction Mapping; Sequence Homology, Amino Acid

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ARATH:KN14E

enables

GO:0005516: calmodulin binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:KN14E

enables

GO:0008017: microtubule binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:KN14E

involved_in

GO:0010091: trichome branching

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:KN14E

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:KN14E

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9ZPX9

F

Seeded From UniProt

complete

ARATH:KIC

enables

GO:0005509: calcium ion binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:KIC

involved_in

GO:0010091: trichome branching

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:KIC

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9FHN8

F

Seeded From UniProt

complete


See also

References

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