title: A genomic toolkit to investigate kinesin and myosin motor function in cells.
abstract: 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.
authors: Max-Planck Institute for Cell Biology and Genetics (CBG), Dresden 01307, Germany. maliga@mpi-cbg.de
authors: Zoltan
authors: Z
authors: Maliga
authors: Magno
authors: M
authors: Junqueira
authors: Yusuke
authors: Y
authors: Toyoda
authors: Andreas
authors: A
authors: Ettinger
authors: Felipe
authors: F
authors: Mora-Bermúdez
authors: Robin W
authors: RW
authors: Klemm
authors: Andrej
authors: A
authors: Vasilj
authors: Elaine
authors: E
authors: Guhr
authors: Itziar
authors: I
authors: Ibarlucea-Benitez
authors: Ina
authors: I
authors: Poser
authors: Ezio
authors: E
authors: Bonifacio
authors: Wieland B
authors: WB
authors: Huttner
authors: Andrej
authors: A
authors: Shevchenko
authors: Anthony A
authors: AA
authors: Hyman
mesh_terms: D000818:Animals; D001692:Biological Transport; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002465:Cell Movement / Q000502:physiology*; D018385:Centrosome / Q000378:metabolism; D002846:Chromatography, Affinity; D022202:Chromosomes, Artificial, Bacterial; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D023281:Genomics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules; D008938:Mitosis / Q000502:physiology; D009218:Myosins / Q000235:genetics / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D055503:Protein Multimerization; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D019076:Transgenes / Q000235:genetics
chemical_list: D015415:Biomarkers; C502822:Cep170 protein, human; C092935:KIF23 protein, human; D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C541430:MKLP1 protein, mouse; D009218:Myosins; D016547:Kinesins