The Calmodulin-binding Domain from a Plant Kinesin Functions as a Modular Domain in Conferring Ca2+-Calmodulin Regulation to Animal Plus- and Minus-end Kinesins

• Published in: The Journal of biological chemistry Vol. 277; no. 50; pp. 48058 - 48065
• Main Authors: Reddy, Vaka S, Reddy, Anireddy S N
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 13.12.2002
• Subjects: Animals; Base Sequence; Binding Sites; Calcium - metabolism; Calmodulin - isolation & purification; Calmodulin - metabolism; DNA Primers; Drosophila; Kinesin - chemistry; Kinesin - isolation & purification; Kinesin - metabolism; Plants - metabolism; Space life sciences; Non-programmatic
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M205459200

Plant kinesin-like calmodulin-binding protein (KCBP) is a novel member of the kinesin superfamily that interacts with calmodulin (CaM) via its CaM-binding domain (CBD). Activated CaM (Ca 2+ -CaM) has been shown to inhibit KCBP interaction with microtubules (MTs) thereby abolishing its motor- and MT-dependent ATPase activities. To test whether the fusion of CBD to non-CaM-binding kinesins confers Ca 2+ -CaM regulation, we fused the CBD of KCBP to the N or C terminus of a minus-end (non-claret disjunction) or C terminus of a plus-end ( Drosophila kinesin) motor. Purified chimeric kinesins bound CaM in a Ca 2+ -dependent manner whereas non-claret disjunction, Drosophila kinesin, and KCBP that lack a CBD did not. As in the case of KCBP with CBD, the interaction of chimeric motors with MTs, as well as their MT-stimulated ATPase activity, was inhibited by Ca 2+ -CaM. The presence of a spacer between the motor and CBD did not alter Ca 2+ -CaM regulation. However, KCBP interaction with MTs and its MT-stimulated ATPase activity were not inhibited when the motor domain and CBD were added separately, suggesting that Ca 2+ -CaM regulation of CaM-binding motors occurs only when the CBD is attached to the motor domain. These results show that the fusion of the CBD to animal motors confers Ca 2+ -CaM regulation and suggest that the CBD functions as a modular domain in disrupting motor-MT interaction. Our data also support the hypothesis that CaM-binding kinesins may have evolved by addition of a CBD to a kinesin motor domain.