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Queens University

1. Delorme, Caroline. Structural and Functional Characterization of a Novel Heterodimeric Kinesin in Candida albicans .

Degree: Biochemistry, 2012, Queens University

Kinesins are molecular motors that transport intracellular cargos along microtubules (MTs) and influence the organization and dynamics of the MT cytoskeleton. Their force-generating functions arise from conformational changes in their motor domain as ATP is bound and hydrolyzed, and products are released. In the budding yeast Saccharomyces cerevisiae, the Kar3 kinesin forms heterodimers with one of two non-catalytic kinesin-like proteins, Cik1 and Vik1, which lack the ability to bind ATP, and yet they retain the capacity to bind MTs. Cik1 and Vik1 also influence and respond to the MT-binding and nucleotide states of Kar3, and differentially regulate the functions of Kar3 during yeast mating and mitosis. The mechanism by which Kar3/Cik1 and Kar3/Vik1 dimers operate remains unknown, but has important implications for understanding mechanical coordination between subunits of motor complexes that traverse cytoskeletal tracks. In this study, we show that the opportunistic human fungal pathogen Candida albicans (Ca) harbors a single version of this unique form of heterodimeric kinesin and we present the first in vitro characterization of this motor. Like its budding yeast counterpart, the Vik1-like subunit binds directly to MTs and strengthens the MT-binding affinity of the heterodimer. However, in contrast to ScKar3/Cik1 and ScKar3/Vik1, CaKar3/Vik1 exhibits weaker overall MT-binding affinity and lower ATPase activity. Preliminary investigations using a multiple motor motility assay indicate CaKar3/Vik1 may not be motile. Using a maltose binding protein tagging system, we determined the X-ray crystal structure of the CaKar3 motor domain and observed notable differences in its nucleotide-binding pocket relative to ScKar3 that appear to represent a previously unobserved state of the active site. Together, these studies broaden our knowledge of novel kinesin motor assemblies and shed new light on structurally dynamic regions of Kar3/Vik1-like motor complexes that help mediate mechanical coordination of its subunits.

Subjects/Keywords: Filamentous Fungus ; Kinesin-14 ; Kar3 ; Candida Albicans ; Switch Elements ; Maltose Binding Protein ; Microtubule Motor Protein ; ATPase activity

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Delorme, C. (2012). Structural and Functional Characterization of a Novel Heterodimeric Kinesin in Candida albicans . (Thesis). Queens University. Retrieved from http://hdl.handle.net/1974/7023

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Delorme, Caroline. “Structural and Functional Characterization of a Novel Heterodimeric Kinesin in Candida albicans .” 2012. Thesis, Queens University. Accessed November 27, 2020. http://hdl.handle.net/1974/7023.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Delorme, Caroline. “Structural and Functional Characterization of a Novel Heterodimeric Kinesin in Candida albicans .” 2012. Web. 27 Nov 2020.

Vancouver:

Delorme C. Structural and Functional Characterization of a Novel Heterodimeric Kinesin in Candida albicans . [Internet] [Thesis]. Queens University; 2012. [cited 2020 Nov 27]. Available from: http://hdl.handle.net/1974/7023.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Delorme C. Structural and Functional Characterization of a Novel Heterodimeric Kinesin in Candida albicans . [Thesis]. Queens University; 2012. Available from: http://hdl.handle.net/1974/7023

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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