Journal Article

Novel asymmetrically localizing components of human centrosomes identified by complementary proteomics methods

Vanselow, Katja, Hyman, Anthony A, Lundberg, Emma, Uhlen, Mathias, Poser, Ina, Bennetzen, Martin, Skogs, Marie, Toyoda, Yusuke, Falkenby, Lasse G, Jakobsen, Lis, Westendorf, Jens, Andersen, Jens S, Nigg, Erich A

2011

Overview

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. Organellar proteomics revealed a surprising complexity of centrosome composition. New combinatorial approaches now further extend the list of centrosome proteins, but also begin to elucidate their dynamics and differential localization.

• Publisher: John Wiley & Sons, Ltd,Nature Publishing Group UK,Springer Nature B.V,Nature Publishing Group
• Subject: Biochemistry / Biokemi / Centrioles - chemistry / Centrioles - metabolism / centrosome / Centrosome - chemistry / Centrosome - metabolism / Chemistry / Cilia - metabolism / Clinical genetics / Dermatologi och venerologi, klinisk genetik, invärtesmedicin / Dermatology and venerology,clinical genetics, internal medicine / EMBO05 / EMBO06 / EMBO43 / Gene loci / HeLa Cells / Humans / Kemi / Klinisk genetik / Mass Spectrometry / mass spectrometry-based proteomics / MEDICIN / MEDICINE / Microscopy, Fluorescence / Molecular biology / Molekylärbiologi / mother and daughter centriole / Organelles / protein turnover / Proteins / Proteins - chemistry / Proteins - metabolism / Proteomics / SILAC