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Volume 271, Number 32, Issue of August 9, 1996 pp. 19358-19366
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Brain Cytoplasmic and Flagellar Outer Arm Dyneins Share a Highly Conserved M_r 8,000 Light Chain

(Received for publication, December 11, 1995, and in revised form, April 15, 1996)

Stephen M. King , Elisa Barbarese ^¶ , James F. Dillman III^'' , Ramila S. Patel-King , John H. Carson and K. Kevin Pfister ^''

From the Departments of  Biochemistry and ^¶ Neurology and the  Center for Biomedical Imaging Technology, University of Connecticut Health Center, Farmington, Connecticut 06032-3305 and the ^'' Department of Cell Biology, University of Virginia Health Science Center, Charlottesville, Virginia 22908-0439

Sequence comparisons with the M_r 8,000 light chain from Chlamydomonas outer arm dynein revealed the presence of highly conserved homologues (up to 90% identity) in the expressed sequence tag data base (King, S. M. & Patel-King, R. S. (1995a) J. Biol. Chem. 270, 11445-11452). Several of these homologous sequences were derived from organisms and/or tissues that lack motile cilia/flagella, suggesting that these proteins may function in the cytoplasm. In Drosophila, lack of the homologous protein results in embryonic lethality (Dick, T., Ray, K., Salz, H. K. & Chia, W. (1996) Mol. Cell. Biol., 16, 1966-1977). Fractionation of mammalian brain homogenates reveals three distinct cytosolic pools of the homologous protein, one of which specifically copurifies with cytoplasmic dynein following both ATP-sensitive microtubule affinity/sucrose density gradient centrifugation and immunoprecipitation with a monoclonal antibody specific for the 74-kDa intermediate chain (IC74). Quantitative densitometry indicates that there is one copy of the M_r 8,000 polypeptide per IC74. Dual channel confocal immunofluorescent microscopy revealed that the M_r 8,000 protein is significantly colocalized with cytoplasmic dynein but not with kinesin in punctate structures (many of which are associated with microtubules) within mammalian oligodendrocytes. Thus, it appears that flagellar outer arm and brain cytoplasmic dyneins share a highly conserved light chain polypeptide that, at least in Drosophila, is essential for viability.

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