Nucleolar and spindle-associated protein 1 (NUSAP1) interacts with a SUMO E3 ligase complex during chromosome segregation

• Published in: The Journal of biological chemistry Vol. 292; no. 42; pp. 17178 - 17189
• Main Authors: Mills, Christine A., Suzuki, Aussie, Arceci, Anthony, Mo, Jin Yao, Duncan, Alex, Salmon, Edward D., Emanuele, Michael J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.10.2017; American Society for Biochemistry and Molecular Biology
• Subjects: Cell Biology; cell cycle; cell division; Chromosome Segregation - drug effects; Chromosome Segregation - physiology; GTPase-Activating Proteins - genetics; GTPase-Activating Proteins - metabolism; HeLa Cells; Humans; microtubule; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; Microtubules - genetics; Microtubules - metabolism; mitosis; mitotic spindle; Molecular Chaperones - genetics; Molecular Chaperones - metabolism; Nuclear Pore Complex Proteins - genetics; Nuclear Pore Complex Proteins - metabolism; Paclitaxel - pharmacology; Protein Domains; small ubiquitin-like modifier (SUMO); Spindle Apparatus - genetics; Spindle Apparatus - metabolism; sumoylation; ubiquitin; Ubiquitin-Conjugating Enzymes - genetics; Ubiquitin-Conjugating Enzymes - metabolism; microtubule; sumoylation; cell cycle; cell division; small ubiquitin-like modifier (SUMO); mitosis; ubiquitin; mitotic spindle
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M117.796045

The mitotic spindle is composed of dynamic microtubules and associated proteins that together direct chromosome movement during mitosis. The spindle plays a vital role in accurate chromosome segregation fidelity and is a therapeutic target in cancer. Nevertheless, the molecular mechanisms by which many spindle-associated proteins function remains unknown. The nucleolar and spindle-associated protein NUSAP1 is a microtubule-binding protein implicated in spindle stability and chromosome segregation. We show here that NUSAP1 localizes to dynamic spindle microtubules in a unique chromosome-centric pattern, in the vicinity of overlapping microtubules, during metaphase and anaphase of mitosis. Mass spectrometry-based analysis of endogenous NUSAP1 interacting proteins uncovered a cell cycle-regulated interaction between the RanBP2–RanGAP1–UBC9 SUMO E3 ligase complex and NUSAP1. Like NUSAP1 depletion, RanBP2 depletion impaired the response of cells to the microtubule poison Taxol. NUSAP1 contains a conserved SAP domain (SAF-A/B, Acinus, and PIAS). SAP domains are common among many other SUMO E3s, and are implicated in substrate recognition and ligase activity. We speculate that NUSAP1 contributes to accurate chromosome segregation by acting as a co-factor for RanBP2–RanGAP1–UBC9 during cell division.