A Dynamic Protein Interaction Landscape of the Human Centrosome-Cilium Interface

Abstract

Summary Centrioles coordinate the primary microtubule organizing center of the cell and template the formation of cilia, thereby operating at a nexus of critical cellular functions. Here we use proximity-dependent biotinylation (BioID) to map the centrosome-cilium interface; with 58 bait proteins we generate a protein topology network comprising >7000 interactions. Analysis of interaction profiles coupled with high resolution phenotypic profiling implicates a number of new protein modules in centriole duplication, ciliogenesis and centriolar satellite biogenesis, and highlights extensive interplay between these processes. By monitoring dynamic changes in the centrosome-cilium protein interaction landscape during ciliogenesis, we also identify satellite proteins that support cilia formation. Systematic profiling of proximity interactions combined with functional analysis thus provides a rich resource for better understanding human centrosome and cilia biology. Similar strategies may be applied to other complex biological structures or pathways.

A Versatile Lentiviral Delivery Toolkit for Proximity-dependent Biotinylation in Diverse Cell Types

“…3). Consistent with previous studies (20,59), the biological replicates in Flp-In T-REx HeLa lines were highly similar, with R 2 =0.99 across all prey proteins identified and R 2 =0.96 when only the high-confidence preys were considered (Fig. 3A).…”

The centriolar satellite protein CCDC66 interacts with CEP290 and functions in cilium formation and trafficking

“…Among these hits is the retinal degeneration gene CCDC66 (Dekomien et al, 2010;Gerding et al, 2011). CCDC66 has been reported to localize to centriolar satellites (Gupta et al, 2015) and its mRNA has been found to interact with microtubules in a screen for microtubule-binding transcripts in Xenopus laevis egg extracts (Sharp et al, 2011). CCDC66 was also previously identified a proximity partner of the canonical satellite protein PCM1 (Gupta et al, 2015), but not for CCDC14 and KIAA0753, which regulate centriole duplication (Firat-Karalar and Stearns, 2015).…”

“…CCDC66 has been reported to localize to centriolar satellites (Gupta et al, 2015) and its mRNA has been found to interact with microtubules in a screen for microtubule-binding transcripts in Xenopus laevis egg extracts (Sharp et al, 2011). CCDC66 was also previously identified a proximity partner of the canonical satellite protein PCM1 (Gupta et al, 2015), but not for CCDC14 and KIAA0753, which regulate centriole duplication (Firat-Karalar and Stearns, 2015). Based on its proximity to regulators of ciliogenesis, disease link and cellular localization, we chose CCDC66 for further study.…”

“…The interactors identified by this map include both known satellite proteins and previously uncharacterized proteins (Firat-Karalar et al, 2014). Among these proteins, we chose CCDC66, for further study, because it has been previously implicated in microtubule-mediated functions, including spindle pole and centriolar satellite organization (Gupta et al, 2015;Sharp et al, 2011), and its link to retinal degeneration (Dekomien et al, 2010;Gerding et al, 2011). A homozygous mutation in the CCDC66 gene was shown to cause autosomal recessively inherited, generalized progressive retinal atrophy, the canine counterpart of Retinitis pigmentosa (Dekomien et al, 2010).…”

Molecular characterization, sequence analysis and tissue expression of a porcine gene –MOSPD2

“…Gene-environment analysis reported that MOSPD2 may be related to the obesity traits among postmenopausal African-American and Hispanic women [4]. This gene is also involved in the dynamic protein interaction of the human centrosome-cilium interface [5].…”