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A more recent version of this article appeared on September 1, 2007
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Submitted on October 13, 2006
Revised on July 3, 2007
Accepted on July 5, 2007
*Department of Molecular, Microbial, and Structural Biology, University of Connecticut Health Center, Farmington, CT 06030;
Kobe Advanced ICT Research Center, National Institute of Information and Communications Technology, 588-2 Iwaoka, Nishi-ku, Kobe 651-2492, Japan
Monitoring Editor: Erika Holzbaur
We have previously shown that Ca2+ directly activates ATP-sensitive microtubule binding by a Chlamydomonas outer arm dynein subparticle containing the
and
heavy chains. The
heavy chain-associated LC4 light chain is a member of the calmodulin family and binds 1-2 Ca2+ with KCa = 3 x 10-5 M in vitro, suggesting it may act as a Ca2+-sensor for outer arm dynein. Here we investigate interactions between the LC4 light chain and
heavy chain. Two IQ consensus motifs for binding calmodulin-like proteins are located within the stem domain of the
heavy chain. In vitro experiments indicate that LC4 undergoes a Ca2+-dependent interaction with the IQ motif domain while remaining tethered to the HC. LC4 also moves into close proximity of the IC IC1 in the presence of Ca2+. The sedimentation profile of the
heavy chain subunit changed subtly upon Ca2+ addition suggesting that the entire complex had become more compact, and electron microscopy of the isolated
subunit revealed a distinct alteration in conformation of the N-terminal stem in response to Ca2+ addition. We propose that Ca2+-dependent conformational change of LC4 has a direct effect on the stem domain of the
heavy chain which eventually leads to alterations in mechanochemical interactions between microtubules and the motor domain(s) of the outer dynein arm.
Present address: Department of Molecular Biology and Biochemistry, Wesleyan University, Middletown, CT.
Address correspondence to:
Stephen M. King (king{at}neuron.uchc.edu)
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