title: Dissecting the domain structure of Cdc4p, a myosin essential light chain involved in Schizosaccharomyces pombe cytokinesis.
abstract: Cytokinesis is the process by which one cell divides into two. Key in the cytokinetic mechanism of Schizosaccharomyces pombe is the contractile ring myosin, which consists of two heavy chains (Myo2p), two essential light chains (Cdc4p), and two regulatory light chains (Rlc1p). Cdc4p is a dumbbell-shaped EF-hand protein composed of N- and C-terminal domains separated by a flexible linker. The properties of these two domains are of particular interest because each is hypothesized to have independent functions in binding different components of the cytokinesis machinery. To help define these properties, we used NMR spectroscopy to compare the structure, stability, and dynamics of the isolated N- and C-terminal domains with one another and with native Cdc4p. On the basis of invariant chemical shifts, the N-domain retains the same structure in isolation as in the context of the full-length Cdc4p, whereas the C-domain appears markedly perturbed. This perturbation results from intramolecular binding of the residual linker sequence at the N-terminus of the C-domain in a mode similar to that used by native Cdc4p to associate with target polypeptide sequences. NMR relaxation, thermal denaturation, and amide hydrogen exchange experiments also indicate that the C-domain is less stable and more dynamic than the N-domain, both in isolation and in the full-length protein. We hypothesize that these properties reflect a conformational plasticity of the C-domain, which may allow Cdc4p to interact with several regulatory or contractile ring proteins necessary for cytokinesis.
authors.affiliation: Department of Biochemistry and Molecular Biology, The Michael Smith Laboratories, and The Protein Engineering Network of Centres of Excellence, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors.forename: Eric
authors.initials: E
authors.lastname: Escobar-Cabrera
authors.forename: Meenakshi
authors.initials: M
authors.lastname: Venkatesan
authors.forename: Michel
authors.initials: M
authors.lastname: Desautels
authors.forename: Sean M
authors.initials: SM
authors.lastname: Hemmingsen
authors.forename: Lawrence P
authors.initials: LP
authors.lastname: McIntosh
mesh_terms: D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D048749:Cytokinesis; D003598:Cytoskeletal Proteins; D044783:F-Box Proteins / Q000737:chemistry* / Q000378:metabolism*; D005998:Glycine / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018994:Myosin Light Chains / Q000737:chemistry* / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012568:Schizosaccharomyces / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D013912:Threonine / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D044783:F-Box Proteins; D018994:Myosin Light Chains; D029702:Schizosaccharomyces pombe Proteins; C513274:cdc4 protein, S pombe; D013912:Threonine; D044767:Ubiquitin-Protein Ligases; D005998:Glycine