title: Novel asymmetrically localizing components of human centrosomes identified by complementary proteomics methods.
abstract: Centrosomes in animal cells are dynamic organelles with a proteinaceous matrix of pericentriolar material assembled around a pair of centrioles. They organize the microtubule cytoskeleton and the mitotic spindle apparatus. Mature centrioles are essential for biogenesis of primary cilia that mediate key signalling events. Despite recent advances, the molecular basis for the plethora of processes coordinated by centrosomes is not fully understood. We have combined protein identification and localization, using PCP-SILAC mass spectrometry, BAC transgeneOmics, and antibodies to define the constituents of human centrosomes. From a background of non-specific proteins, we distinguished 126 known and 40 candidate centrosomal proteins, of which 22 were confirmed as novel components. An antibody screen covering 4000 genes revealed an additional 113 candidates. We illustrate the power of our methods by identifying a novel set of five proteins preferentially associated with mother or daughter centrioles, comprising genes implicated in cell polarity. Pulsed labelling demonstrates a remarkable variation in the stability of centrosomal protein complexes. These spatiotemporal proteomics data provide leads to the further functional characterization of centrosomal proteins.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
authors.forename: Lis
authors.initials: L
authors.lastname: Jakobsen
authors.forename: Katja
authors.initials: K
authors.lastname: Vanselow
authors.forename: Marie
authors.initials: M
authors.lastname: Skogs
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Toyoda
authors.forename: Emma
authors.initials: E
authors.lastname: Lundberg
authors.forename: Ina
authors.initials: I
authors.lastname: Poser
authors.forename: Lasse G
authors.initials: LG
authors.lastname: Falkenby
authors.forename: Martin
authors.initials: M
authors.lastname: Bennetzen
authors.forename: Jens
authors.initials: J
authors.lastname: Westendorf
authors.forename: Erich A
authors.initials: EA
authors.lastname: Nigg
authors.forename: Mathias
authors.initials: M
authors.lastname: Uhlen
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Hyman
authors.forename: Jens S
authors.initials: JS
authors.lastname: Andersen
mesh_terms: D002502:Centrioles / Q000737:chemistry / Q000378:metabolism; D018385:Centrosome / Q000737:chemistry / Q000378:metabolism*; D002923:Cilia / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D015388:Organelles; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics
chemical_list: D011506:Proteins