Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation Sites
Nucleolar and Spindle-Associated Protein 1 (NuSAP1) is an important mitotic regulator, implicated in control of mitotic microtubule stability and chromosome segregation. NuSAP1 regulates these processes by interacting with several protein partners. Its abundance, activity and interactions are theref...
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| Format: | Article |
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MDPI AG
2023-10-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/21/2545 |
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| author | Michela Damizia Ludovica Altieri Vincenzo Costanzo Patrizia Lavia |
| author_facet | Michela Damizia Ludovica Altieri Vincenzo Costanzo Patrizia Lavia |
| author_sort | Michela Damizia |
| collection | DOAJ |
| description | Nucleolar and Spindle-Associated Protein 1 (NuSAP1) is an important mitotic regulator, implicated in control of mitotic microtubule stability and chromosome segregation. NuSAP1 regulates these processes by interacting with several protein partners. Its abundance, activity and interactions are therefore tightly regulated during mitosis. Protein conjugation with SUMO (Small Ubiquitin-like MOdifier peptide) is a reversible post-translational modification that modulates rapid changes in the structure, interaction(s) and localization of proteins. NuSAP1 was previously found to interact with RANBP2, a nucleoporin with SUMO ligase and SUMO-stabilizing activity, but how this interaction affects NuSAP1 activity has remained elusive. Here, we show that NuSAP1 interacts with RANBP2 and forms proximity ligation products with SUMO2/3 peptides in a RANBP2-dependent manner at key mitotic sites. A bioinformatic search identified two putative SUMO consensus sites in NuSAP1, within the DNA-binding and the microtubule-binding domains, respectively. Site-specific mutagenesis, and mitotic phenotyping in cell lines expressing each NuSAP1 mutant version, revealed selective roles of each individual site in control of NuSAP1 localization and in generation of specific mitotic defects and distinct fates in daughter cells. These results identify therefore two new regulatory sites for NuSAP1 functions and implicate RANBP2 in control of NuSAP1 activity. |
| first_indexed | 2024-03-11T11:32:20Z |
| format | Article |
| id | doaj.art-14bc738d85604e50b6ded09ca9362157 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T11:32:20Z |
| publishDate | 2023-10-01 |
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| spelling | doaj.art-14bc738d85604e50b6ded09ca93621572023-11-10T15:00:43ZengMDPI AGCells2073-44092023-10-011221254510.3390/cells12212545Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation SitesMichela Damizia0Ludovica Altieri1Vincenzo Costanzo2Patrizia Lavia3Institute of Molecular Biology and Pathology (IBPM), CNR National Research Council of Italy, 00185 Rome, ItalyInstitute of Molecular Biology and Pathology (IBPM), CNR National Research Council of Italy, 00185 Rome, ItalyInstitute of Molecular Biology and Pathology (IBPM), CNR National Research Council of Italy, 00185 Rome, ItalyInstitute of Molecular Biology and Pathology (IBPM), CNR National Research Council of Italy, 00185 Rome, ItalyNucleolar and Spindle-Associated Protein 1 (NuSAP1) is an important mitotic regulator, implicated in control of mitotic microtubule stability and chromosome segregation. NuSAP1 regulates these processes by interacting with several protein partners. Its abundance, activity and interactions are therefore tightly regulated during mitosis. Protein conjugation with SUMO (Small Ubiquitin-like MOdifier peptide) is a reversible post-translational modification that modulates rapid changes in the structure, interaction(s) and localization of proteins. NuSAP1 was previously found to interact with RANBP2, a nucleoporin with SUMO ligase and SUMO-stabilizing activity, but how this interaction affects NuSAP1 activity has remained elusive. Here, we show that NuSAP1 interacts with RANBP2 and forms proximity ligation products with SUMO2/3 peptides in a RANBP2-dependent manner at key mitotic sites. A bioinformatic search identified two putative SUMO consensus sites in NuSAP1, within the DNA-binding and the microtubule-binding domains, respectively. Site-specific mutagenesis, and mitotic phenotyping in cell lines expressing each NuSAP1 mutant version, revealed selective roles of each individual site in control of NuSAP1 localization and in generation of specific mitotic defects and distinct fates in daughter cells. These results identify therefore two new regulatory sites for NuSAP1 functions and implicate RANBP2 in control of NuSAP1 activity.https://www.mdpi.com/2073-4409/12/21/2545NuSAP1RANBP2mitosismicrotubuleskinetochoresSUMOylation |
| spellingShingle | Michela Damizia Ludovica Altieri Vincenzo Costanzo Patrizia Lavia Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation Sites Cells NuSAP1 RANBP2 mitosis microtubules kinetochores SUMOylation |
| title | Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation Sites |
| title_full | Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation Sites |
| title_fullStr | Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation Sites |
| title_full_unstemmed | Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation Sites |
| title_short | Distinct Mitotic Functions of Nucleolar and Spindle-Associated Protein 1 (NuSAP1) Are Controlled by Two Consensus SUMOylation Sites |
| title_sort | distinct mitotic functions of nucleolar and spindle associated protein 1 nusap1 are controlled by two consensus sumoylation sites |
| topic | NuSAP1 RANBP2 mitosis microtubules kinetochores SUMOylation |
| url | https://www.mdpi.com/2073-4409/12/21/2545 |
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