Dresden 2011 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 33: Biological Machines \& Motor Proteins
BP 33.7: Vortrag
Freitag, 18. März 2011, 12:15–12:30, ZEU 250
Kinesin-3 (UNC-104) can act as a dimeric motor during axonal transport C. elegans neurons in vivo — •Volker Christoph Henschel1, Alessandro Esposito2, Christoph Friedrich Schmidt1, Fred Sylvester Wouters3, and Dieter Robert Klopfenstein1 — 1Drittes Physikalisches Institut, Biophysics, Georg-August-University Göttingen, Göttingen, Germany — 2MRC Cancer Cell Unit, Hutchison/MRC Research Centre, Cambridge, UK — 3Laboratory of Cellular and Molecular Systems, Department of Neuro- and Sensory Physiology, Georg-August-University Göttingen, Göttingen Germany
Monomeric Kinesin-3 (UNC-104) is responsible for the transport of presynaptic vesicles to synaptic termini in C. elegans. To investigate the role of the endogenous coiled-coils, we introduced point mutations in the motors coiled-coil region in the neck promoting either dimer formation of Kinesin-3 or reducing the likelihood of dimerization. We verify dimerization by cross-linking of purified truncated motors in vitro. We show by live in vivo imaging, that reducing dimerization of Kinesin-3 leads to decreased vesicle transport velocities and affects the control of muscle contraction. C.elegans with reduced dimerization properties exhibit a 45% reduction in anterograde velocity. Additionally, severe motility and a significant egg laying defect are observed. To assess dimer formation in vivo we combine Foerster Resonance Energy Transfer (FRET) and anisotropy imaging with spinning-disc laser confocal microscopy. Our data suggest a direct link between dimerization status and transport velocities.