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J. Biol. Chem., Vol. 276, Issue 17, 14366-14373, April 27, 2001
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From the Department of Biochemistry, University of Connecticut
Health Center, Farmington, Connecticut 06030-3305
The Tctex1/Tctex2 family of dynein light chains
associates with the intermediate chains at the base of the soluble
dynein particle. These components are essential for dynein assembly and participate in specific motor-cargo interactions. To further
address the role of these light chains in dynein activity, the
structural and biochemical properties of several members of this
polypeptide class were examined. Gel filtration chromatography and
native gel electrophoresis indicate that recombinant
Chlamydomonas flagellar Tctex1 exists as a dimer in
solution. Furthermore, yeast two-hybrid analysis suggests that this
association also occurs in vivo. In contrast, both murine
and Chlamydomonas Tctex2 are monomeric. To investigate
protein-protein interactions involving these light chains, outer arm
dynein from Chlamydomonas flagella was cross-linked using
dimethylpimelimidate. Immunoblot analysis of the resulting products revealed the interaction of LC2 (Tctex2) with LC6, which is
closely related to the highly conserved LC8 protein found in many
enzyme systems, including dynein. Northern dot blot analysis demonstrated that Tctex1/Tctex2 family light chains are differentially expressed both in a tissue-specific and developmentally regulated manner in humans. These data provide further support for the existence of functionally distinct populations of cytoplasmic dynein with differing light chain content.
The Tctex1/Tctex2 Class of Dynein Light Chains
DIMERIZATION, DIFFERENTIAL EXPRESSION, AND INTERACTION WITH THE
LC8 PROTEIN FAMILY*
,
*
This study was supported in part by Grant GM51293 from the
National Institutes of Health and by the Heritage Affiliate of the
American Heart Association.The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Present address: Dept. of Molecular Genetics, The Ohio State
University, Columbus, OH 43210.
§
An investigator of the Patrick and Catherine Weldon Donaghue
Medical Research Foundation. To whom correspondence should be addressed: Dept. of Biochemistry, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030-3305. Tel:
860-679-3347; Fax: 860-679-3408; E-mail: steve@king2.uchc.edu.
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