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A genomic toolkit to investigate kinesin and myosin motor function in cells.
Nat. Cell Biol. 15, 325-334 (2013)
Coordination of multiple kinesin and myosin motors is required for intracellular transport, cell motility and mitosis. However, comprehensive resources that allow systems analysis of the localization and interplay between motors in living cells do not exist. Here, we generated a library of 243 amino- and carboxy-terminally tagged mouse and human bacterial artificial chromosome transgenes to establish 227 stably transfected HeLa cell lines, 15 mouse embryonic stem cell lines and 1 transgenic mouse line. The cells were characterized by expression and localization analyses and further investigated by affinity-purification mass spectrometry, identifying 191 candidate protein-protein interactions. We illustrate the power of this resource in two ways. First, by characterizing a network of interactions that targets CEP170 to centrosomes, and second, by showing that kinesin light-chain heterodimers bind conventional kinesin in cells. Our work provides a set of validated resources and candidate molecular pathways to investigate motor protein function across cell lineages.
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Publication type Article: Journal article
Document type Scientific Article
ISSN (print) / ISBN 1465-7392
Journal Nature Cell Biology
Quellenangaben Volume: 15, Issue: 3, Pages: 325-334
Publisher Nature Publishing Group
Reviewing status Peer reviewed
Institute(s) Institute for Pancreatic Beta Cell Research (IPI)