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A Novel Kinesin-Like Protein with a Calmodulin-Binding DomainCalcium regulates diverse developmental processes in plants through the action of calmodulin. A cDNA expression library from developing anthers of tobacco was screened with S-35-labeled calmodulin to isolate cDNAs encoding calmodulin-binding proteins. Among several clones isolated, a kinesin-like gene (TCK1) that encodes a calmodulin-binding kinesin-like protein was obtained. The TCK1 cDNA encodes a protein with 1265 amino acid residues. Its structural features are very similar to those of known kinesin heavy chains and kinesin-like proteins from plants and animals, with one distinct exception. Unlike other known kinesin-like proteins, TCK1 contains a calmodulin-binding domain which distinguishes it from all other known kinesin genes. Escherichia coli-expressed TCK1 binds calmodulin in a Ca(2+)-dependent manner. In addition to the presence of a calmodulin-binding domain at the carboxyl terminal, it also has a leucine zipper motif in the stalk region. The amino acid sequence at the carboxyl terminal of TCK1 has striking homology with the mechanochemical motor domain of kinesins. The motor domain has ATPase activity that is stimulated by microtubules. Southern blot analysis revealed that TCK1 is coded by a single gene. Expression studies indicated that TCKI is expressed in all of the tissues tested. Its expression is highest in the stigma and anther, especially during the early stages of anther development. Our results suggest that Ca(2+)/calmodulin may play an important role in the function of this microtubule-associated motor protein and may be involved in the regulation of microtubule-based intracellular transport.
Document ID
19970021386
Acquisition Source
Kennedy Space Center
Document Type
Reprint (Version printed in journal)
External Source(s)
doi:10.1007/BF00020609
Authors
Wang, W. (Washington State Univ. Pullman, WA United States)
Takezawa, D. (Washington State Univ. Pullman, WA United States)
Narasimhulu, S. B. (Colorado State Univ. Fort Collins, CO United States)
Reddy, A. S. N. (Colorado State Univ. Fort Collins, CO United States)
Poovaiah, B. W. (Washington State Univ. Pullman, WA United States)
Date Acquired
September 6, 2013
Publication Date
January 1, 1996
Publication Information
Publication: Plant Molecular Biology
Publisher: Kluwer Academic Publishers
Issue: 31
Subject Category
Life Sciences (General)
Report/Patent Number
NAS 1.26:204645
NASA-CR-204645
Accession Number
97N22378
Funding Number(s)
CONTRACT_GRANT: NAG10-0061
CONTRACT_GRANT: NSF DCB-91-4586
Distribution Limits
Public
Copyright
Public Use Permitted.

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