Identification of a Microtubule-binding Domain in a Cytoplasmic Dynein Heavy Chain

As a molecular motor, dynein must coordinate ATP hydrolysis with conformational changes that lead to processive interactions with a microtubule and generate force. To understand how these processes occur, we have begun to map functional domains of a dynein heavy chain from Dictyostelium. The carboxy...

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Bibliographic Details
Published in:The Journal of biological chemistry 1997-08, Vol.272 (32), p.19714-19718
Main Author: Koonce, Michael P.
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - metabolism
Animals
BINDING SITES
Cytoplasm - enzymology
DELETIONS
DICTYOSTELIUM
DICTYOSTELIUM DISCOIDEUM
DYNEIN ATPASE
Dyneins - chemistry
Dyneins - genetics
Dyneins - metabolism
GENE
GENES
HIDROLASAS
HYDROLASE
HYDROLASES
Hydrolysis
MICROTUBULE
MICROTUBULES
Microtubules - metabolism
MICROTUBULOS
MOLECULAR CONFORMATION
Molecular Sequence Data
Open Reading Frames
PARTIAL DELETIONS
PEPTIDE
PEPTIDES
PEPTIDOS
POLYPEPTIDES
Protein Binding
Protein Biosynthesis
Protein Conformation
Protein Folding
PYROPHOSPHATASES
SECONDARY STRUCTURE
Sequence Deletion
Transcription, Genetic
Ultraviolet Rays
Vanadates - metabolism
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Summary:As a molecular motor, dynein must coordinate ATP hydrolysis with conformational changes that lead to processive interactions with a microtubule and generate force. To understand how these processes occur, we have begun to map functional domains of a dynein heavy chain from Dictyostelium. The carboxyl-terminal 10-kilobase region of the heavy chain encodes a 380-kDa polypeptide that approximates the globular head domain. Attempts to further truncate this region fail to produce polypeptides that either bind microtubules or UV-vanadate cleave, indicating that the entire 10-kilobase fragment is necessary to produce a properly folded functional dynein head. We have further identified a region just downstream from the fourth P-loop that appears to constitute at least part of the microtubule-binding domain (amino acids 3182–3818). When deleted, the resulting head domain polypeptide no longer binds microtubules; when the excised region is expressed in vitro, it cosediments with added tubulin polymer. This microtubule-binding domain falls within an area of the molecule predicted to form extended α-helices. At least four discrete sites appear to coordinate activities required to bind the tubulin polymer, indicating that the interaction of dynein with microtubules is complex.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.272.32.19714