A Proximity Mapping Journey into the Biology of the Mammalian Centrosome/Cilium Complex

Abstract

The mammalian centrosome/cilium complex is composed of the centrosome, the primary cilium and the centriolar satellites, which together regulate cell polarity, signaling, proliferation and motility in cells and thereby development and homeostasis in organisms. Accordingly, deregulation of its structure and functions is implicated in various human diseases including cancer, developmental disorders and neurodegenerative diseases. To better understand these disease connections, the molecular underpinnings of the assembly, maintenance and dynamic adaptations of the centrosome/cilium complex need to be uncovered with exquisite detail. Application of proximity-based labeling methods to the centrosome/cilium complex generated spatial and temporal interaction maps for its components and provided key insights into these questions. In this review, we first describe the structure and cell cycle-linked regulation of the centrosome/cilium complex. Next, we explain the inherent biochemical and temporal limitations in probing the structure and function of the centrosome/cilium complex and describe how proximity-based labeling approaches have addressed them. Finally, we explore current insights into the knowledge we gained from the proximity mapping studies as it pertains to centrosome and cilium biogenesis and systematic characterization of the centrosome, cilium and centriolar satellite interactomes.

Proximity labeling at non-centrosomal microtubule-organizing centers reveals VAB-10B and WDR-62 as distinct microtubule regulators

“…Unlike ncMTOCs, essential centrosome components have been defined over the last two decades, a feat made possible by both genetic and proteomic approaches (Bauer et al, 2016;Arslanhan et al, 2020). Neither approach has been used systematically to identify ncMTOC components, perhaps due to a lack of clear phenotypes to screen for and the difficulty in purifying these enigmatic structures.…”

Aurora Kinase A proximity interactome reveals centriolar satellites as regulators of its function during primary cilium biogenesis

“…To further our understanding of AURKA, spatial AURKA interactomes for its different sub-cellular pools and temporal AURKA interactomes in response to different stimuli should be generated. To this end, biochemical purifications of different cellular compartments can be combined with proteomics approaches that have high temporal resolution such as TurboID-based proximity labeling (Arslanhan et al, 2020, Gingras et al, 2019).…”

“…In this study, we applied the in vivo proximity-dependent biotin identification (BioID) approach to map the proximity interactome of AURKA of asynchronous cells. BioID makes use of the promiscuous E. coli biotin ligase BirA(R118G) (hereafter BirA*), which enables covalent biotinylation of proteins within about 10-20 nm proximity of BirA*-fusions of bait proteins (Arslanhan et al, 2020, Roux et al, 2012). The resulting interactome provides a powerful resource for dissecting the dynamic AURKA sub-complexes that enable its multifaceted regulation and diverse AURKA functions.…”

Centrosome Dynamics and Its Role in Inflammatory Response and Metastatic Process

“…Centrosomes promote the production of bipolar mitotic spindles and supply a matrix of primary cilia in various cell types ( Figure 2 A) [ 21 , 22 ]. In addition to these structural functions, centrosomes and primary cilia have also evolved into essential signaling hubs to build and regulate the sensory/motor characters of metazoans [ 23 , 24 ]. It is increasingly recognized that centrosome is an organizing unit not only for MTs, but also for actin, Golgi apparatus, and signaling molecules coordinating cell migration, cell polarity, and fundamental signal transduction pathways [ 25 , 26 ].…”