A rapid computational approach identifies SPICE1 as an Aurora kinase substrate

Abstract

Aurora kinases play a major role in mitosis by regulating diverse substrates. Defining their critical downstream targets is important in understanding Aurora kinase function. Here we have developed an unbiased computational approach to identify new Aurora kinase substrates based on phosphorylation site clustering, protein localization, protein structure, and species conservation. We validate the microtubule-associated proteins Clasp2, Elys, tubulin tyrosine ligase-like polyglutamylase residues 330–624 and spindle and centriole associated protein 1, residues 549–855 (SPICE1), as Aurora A and B kinases substrates in vitro. We also demonstrate that SPICE1 localization is regulated by Aurora kinases during mitosis. In the absence of Aurora kinase activity, SPICE1 remains at centrioles but does not target to the spindle. Similarly, a nonphosphorylatable SPICE1 mutant no longer localizes to the spindle. Finally, we show that misregulating SPICE1 phosphorylation results in abnormal centriole number, spindle multipolarity, and chromosome alignment defects. Overall, our work indicates that temporal and spatial Aurora kinase–mediated regulation of SPICE1 is important for correct chromosome segregation. In addition, our work provides a database-search tool that enables rapid identification of Aurora kinase substrates.

BUB-1 targets PP2A:B56 to regulate chromosome congression during meiosis I in C. elegans oocytes

“…In this respect, we found that serine 612 in BUB-1 is phosphorylated and this serine is embedded in a sequence (RRL S I) closely resembling the consensus for PP2A:B56 reported in Kruse et al, 2020 . Furthermore, the sequence also fits into the loosely defined Aurora B consensus RRxSφ (where φ is a hydrophobic aa) ( Cheeseman et al, 2002 ; Deretic et al, 2019 ; Kettenbach et al, 2011 ). Therefore, BUB-1 itself could be subjected to an Aurora B-PP2A:B56 balance and it will be very interesting to address whether BUB-1 and other meiotic proteins are regulated by the balance between Aurora B and PP2A:B56.…”

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

“…Since its identification as a mitotic kinase, research on AURKA has extensively focused on its characterization during mitosis (Barr and Gergely, 2007, Nikonova et al, 2013). Multiple low and high throughput conventional approaches were used to identify mitotic AURKA interactors, which fell short in providing mechanistic insight into the emerging non-mitotic functions of AURKA such as cilium assembly and disassembly (Deretic et al, 2019, Kettenbach et al, 2011, Santamaria et al, 2011). In this study, we overcame this limitation by identifying the proximity interactors of AURKA in asynchronous cells using the BioID approach.…”

Phospho‐regulation of mitotic spindle assembly

“…Computational approaches, such as ScanSite (Obenauer, Cantley, & Yaffe, 2003) or PhosphoPredict (Song et al, 2017) have been developed that analyze phosphosites and their motifs and predict potential substrates, such as the prediction and validation of SPICE1 as an Aurora A and Aurora B substrate (Deretic, Kerr, & Welburn, 2019).…”