Sequence and phylogenetic analysis of squid myosin-V: A vesicle motor in nerve cells

Bradley J. Molyneaux, Mary K. Mulcahey, Phillip Stafford, George M. Langford

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We have shown that vesicles in the axoplasm of the squid giant axon move on actin filaments and that movement is inhibited by myosin V-specific antibodies [Tabb et al., 1998]. In the study reported in this article, experiments were performed to clone and sequence the cDNA for squid brain myosin V. Five proteolytic fragments of purified squid brain myosin V were analyzed by direct protein sequencing [Tabb et al., 1998]. Based on this sequence information, degenerate primers were constructed and used to isolate cDNA clones by PCR. Five clones, representing overlapping segments of the gene, were sequenced. The sequence data and the previous biochemical characterization of the molecule support the classification of this vesicle-associated myosin as a member of the class V myosins. Motif analysis of the head, neck, and tail domains revealed that squid MyoV has consensus sequences for all the motifs found in vertebrate members of the myosin V family of motor proteins. A phylogenetic tree was constructed from a sequence alignment by the neighbor- joining method, using Megalign (DNAStar, Madison, WI); the resulting phylogenetic tree showed that squid MyoV is more closely related to vertebrate MyoV (mouse dilute, chicken dilute, rat myr6, and human myo5a) than Drosophila and yeast (myo2, and myo4) myosins V. These new data on the phylogenetic relationships of squid myosin V to vertebrate myosin V strengthens the argument that myosin V functions as a vesicle motor in vertebrate neurons. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish (US)
Pages (from-to)108-115
Number of pages8
JournalCell Motility and the Cytoskeleton
Volume46
Issue number2
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Myosin Type V
Decapodiformes
Sequence Analysis
Neurons
Vertebrates
Clone Cells
Complementary DNA
Overlapping Genes
Sequence Alignment
Protein Sequence Analysis
Consensus Sequence
Brain
Myosins
Actin Cytoskeleton
Drosophila
Axons
Tail
Chickens
Neck
Yeasts

Keywords

  • Actin filaments
  • Axonal transport
  • Giant axon
  • Molecular motor
  • Vesicle/organelle movement

ASJC Scopus subject areas

  • Cell Biology

Cite this

Sequence and phylogenetic analysis of squid myosin-V : A vesicle motor in nerve cells. / Molyneaux, Bradley J.; Mulcahey, Mary K.; Stafford, Phillip; Langford, George M.

In: Cell Motility and the Cytoskeleton, Vol. 46, No. 2, 2000, p. 108-115.

Research output: Contribution to journalArticle

Molyneaux, Bradley J. ; Mulcahey, Mary K. ; Stafford, Phillip ; Langford, George M. / Sequence and phylogenetic analysis of squid myosin-V : A vesicle motor in nerve cells. In: Cell Motility and the Cytoskeleton. 2000 ; Vol. 46, No. 2. pp. 108-115.
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