Proximity mapping of the microtubule plus-end tracking protein SLAIN2 using the BioID approach.

Elif Nur Firat-Karalar

doi:10.3906/biy-2002-12

Abstract

The centrosome is the main microtubule-organizing center of animal cells, which plays key roles in critical cellular processes ranging from cell division to cellular signaling. Accordingly, defects in the structure and function of centrosomes cause various human diseases such as cancer and primary microcephaly. To elucidate the molecular defects underlying these diseases, the biogenesis and functions of the centrosomes have to be fully understood. An essential step towards addressing these questions is the identification and functional dissection of the full repertoire of centrosome proteins. Here, we used high-resolution imaging and showed that the microtubule plus-end tracking protein SLAIN2 localizes to the pericentriolar material at the proximal end of centrioles. To gain insight into its cellular functions and mechanisms, we applied in vivo proximity-dependent biotin identification to SLAIN2 and generated its proximity interaction map. Gene ontology analysis of the SLAIN2 interactome revealed extensive interactions with centriole duplication, ciliogenesis, and microtubule-associated proteins, including previously characterized and uncharacterized interactions. Collectively, our results define SLAIN2 as a component of pericentriolar material and provide an important resource for future studies aimed at elucidating SLAIN2 functions at the centrosome.

Highlights

The centrosome is the main microtubule-organizing center of animal cells, which play skey roles in critical cellular processes ranging from cell division to cellular signaling (Chavali et al, 2014)
SLAIN2 localizes to the centrosome throughout the cell cycle To investigate the cellular functions and mechanisms of SLAIN2 at the centrosome, we examined the localization of endogenous SLAIN2 and a V5-SLAIN2 fusion protein in human osteosarcoma U2OS cells, which are wellcharacterized for their centrosome biogenesis and ideal for visualizing the microtubule network due to their flat morphology
SLAIN2 is a microtubule plus-end tracking protein implicated in cancer

Summary

Introduction

The centrosome is the main microtubule-organizing center of animal cells, which play skey roles in critical cellular processes ranging from cell division to cellular signaling (Chavali et al, 2014). The centrosome consists of two cylindrical microtubule-based structures termed centrioles and associated pericentriolar material, which supports microtubule nucleation, polymerization, and stability (Bettencourt-Dias and Glover, 2007; Luders and Stearns, 2007; Nigg and Raff, 2009). Centrosomes organize the interphase microtubule network, which is required for vesicular trafficking, cell migration and cell shape (Luders and Stearns, 2007). Centrosomes form the mitotic spindle, which mediates equal segregation of genetic material to daughter cells (Nigg and Raff, 2009). In some cycling cells and most quiescent noncycling cells, one of the centrioles forms the basal body that nucleates the microtubule axoneme of the primary cilium.

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