Characterizing the cargo binding and regulatory function of the tail domain in Ncd motor protein

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dc.contributor.authorLonergan, Natalie Elaineen
dc.contributor.committeechairWalker, Richard A.en
dc.contributor.committeememberWong, Eric A.en
dc.contributor.committeememberSible, Jill C.en
dc.contributor.departmentBiologyen
dc.date.accessioned2014-03-14T20:47:06Zen
dc.date.adate2009-11-23en
dc.date.available2014-03-14T20:47:06Zen
dc.date.issued2009-10-09en
dc.date.rdate2009-11-23en
dc.date.sdate2009-10-27en
dc.description.abstractNon-claret disjunctional (Ncd) is a kinesin-14 microtubule motor protein involved in the assembly and stability of meiotic and mitotic spindles in Drosophila oocytes and early embryos, respectively. Ncd functions by cross-linking microtubules through the tail and motor domains. It was originally believed that the role of the Ncd tail domain was to only statically bind microtubules. However, the Ncd tail domain has recently been shown to have properties that stabilize and bundle microtubules, and contribute to the overall motility of the Ncd protein. Continued characterization of the Ncd tail domain is essential to understanding the complete role of Ncd in cell division. This work explored the regulatory function and microtubule binding properties of the Ncd tail domain. Ncd activity is regulated during interphase by nuclear sequestration. GFP-Ncd fusion proteins, containing full length Ncd, individual Ncd domains, or combinations of Ncd domains, were used to identify the presence of a nuclear localization signal (NLS) in the Ncd polypeptide. The nuclear localization of only the GFP fusion proteins containing the Ncd tail sequence indicates that the NLS is contained within the tail domain. Subsequent, experiments performed with GFP fusion proteins containing segments of the tail domain indicate that essential NLS amino acid segments may span the length of the tail domain. Attempts to characterize the microtubule binding properties of the Ncd tail domain, using bacterially expressed MBP-Ncd tail-stalk, were unsuccessful. MBP-Ncd tail-stalk proteins aggregated under binding assay conditions, preventing an accurate determination of the stoichiometric binding relationship between Ncd and the tubulin dimer.en
dc.description.degreeMaster of Scienceen
dc.identifier.otheretd-10272009-084027en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-10272009-084027/en
dc.identifier.urihttp://hdl.handle.net/10919/35511en
dc.publisherVirginia Techen
dc.relation.haspartLonergan_NE_T_2009.pdfen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectnon-claret disjunctional proteinen
dc.subjectnuclear localization signalen
dc.subjectKinesin-14 motor proteinsen
dc.titleCharacterizing the cargo binding and regulatory function of the tail domain in Ncd motor proteinen
dc.typeThesisen
thesis.degree.disciplineBiologyen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.levelmastersen
thesis.degree.nameMaster of Scienceen

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