AMBRA1 phosphorylation by CDK1 and PLK1 regulates mitotic spindle orientation.

Humans Phosphorylation Protein Serine-Threonine Kinases / genetics Spindle Apparatus / metabolism Cell Cycle Proteins / metabolism Mitosis Cell Cycle HeLa Cells CDC2 Protein Kinase / metabolism Adaptor Proteins, Signal Transducing / metabolism

Cell cycle Mitotic kinases Mitotic spindle NUMA1 Phosphorylation

Journal

Cellular and molecular life sciences : CMLS

ISSN: 1420-9071

Titre abrégé: Cell Mol Life Sci

Pays: Switzerland

ID NLM: 9705402

Informations de publication

Date de publication:
16 Aug 2023

Historique:

received: 31 12 2022

accepted: 17 07 2023

revised: 27 06 2023

medline: 17 8 2023

pubmed: 16 8 2023

entrez: 16 8 2023

Statut: epublish

Résumé

AMBRA1 is a crucial factor for nervous system development, and its function has been mainly associated with autophagy. It has been also linked to cell proliferation control, through its ability to regulate c-Myc and D-type cyclins protein levels, thus regulating G1-S transition. However, it remains still unknown whether AMBRA1 is differentially regulated during the cell cycle, and if this pro-autophagy protein exerts a direct role in controlling mitosis too. Here we show that AMBRA1 is phosphorylated during mitosis on multiple sites by CDK1 and PLK1, two mitotic kinases. Moreover, we demonstrate that AMBRA1 phosphorylation at mitosis is required for a proper spindle function and orientation, driven by NUMA1 protein. Indeed, we show that the localization and/or dynamics of NUMA1 are strictly dependent on AMBRA1 presence, phosphorylation and binding ability. Since spindle orientation is critical for tissue morphogenesis and differentiation, our findings could account for an additional role of AMBRA1 in development and cancer ontogenesis.

Identifiants

DOI: 10.1007/s00018-023-04878-6 PMID: 37584777 PMC: PMC10432340

pubmed: 37584777

doi: 10.1007/s00018-023-04878-6

pii: 10.1007/s00018-023-04878-6

pmc: PMC10432340

doi:

Substances chimiques

Protein Serine-Threonine Kinases EC 2.7.11.1

Cell Cycle Proteins 0

CDK1 protein, human EC 2.7.11.22

CDC2 Protein Kinase EC 2.7.11.22

AMBRA1 protein, human 0

Adaptor Proteins, Signal Transducing 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

Pagination

251

Subventions

Organisme : Kræftens Bekæmpelse

ID : R72-A4408

Organisme : Kræftens Bekæmpelse

ID : R146-A9364

Organisme : Novo Nordisk Fonden

ID : 7559

Organisme : Novo Nordisk Fonden

ID : 22544

Organisme : Lundbeckfonden

ID : R233-2016-3360

Organisme : LEO Fondet

ID : LF17024

Organisme : Associazione Italiana per la Ricerca sul Cancro

ID : 23543

Informations de copyright

Références

Fimia GM, Stoykova A, Romagnoli A et al (2007) Ambra1 regulates autophagy and development of the nervous system. Nature 447:1121–1125. https://doi.org/10.1038/nature05925

doi: 10.1038/nature05925 pubmed: 17589504

Cianfanelli V, Fuoco C, Lorente M et al (2015) AMBRA1 links autophagy to cell proliferation and tumorigenesis by promoting c-Myc dephosphorylation and degradation. Nat Cell Biol 17:20–30. https://doi.org/10.1038/ncb3072

doi: 10.1038/ncb3072 pubmed: 25438055

Di Rita A, Strappazzon F (2017) AMBRA1, a novel BH3-like protein: new insights into the AMBRA1–BCL2-family proteins relationship. Int Rev Cell Mol Biol 330:85–113. https://doi.org/10.1016/bs.ircmb.2016.09.002

doi: 10.1016/bs.ircmb.2016.09.002 pubmed: 28215535

Di Rita A, D’Acunzo P, Simula L et al (2018) AMBRA1-mediated mitophagy counteracts oxidative stress and apoptosis induced by neurotoxicity in human neuroblastoma SH-SY5Y cells. Front Cell Neurosci 12:1–11. https://doi.org/10.3389/fncel.2018.00092

doi: 10.3389/fncel.2018.00092

Gu W, Wan D, Qian Q et al (2014) Ambra1 is an essential regulator of autophagy and apoptosis in SW620 cells: pro-survival role of Ambra1. PLoS ONE 9:e90151. https://doi.org/10.1371/journal.pone.0090151

doi: 10.1371/journal.pone.0090151 pubmed: 24587252 pmcid: 3936000

Nazio F, Strappazzon F, Antonioli M et al (2013) mTOR inhibits autophagy by controlling ULK1 ubiquitylation, self-association and function through AMBRA1 and TRAF6. Nat Cell Biol 15:406–416. https://doi.org/10.1038/ncb2708

doi: 10.1038/ncb2708 pubmed: 23524951

Strappazzon F, Di Rita A, Cianfanelli V et al (2016) Prosurvival AMBRA1 turns into a proapoptotic BH3-like protein during mitochondrial apoptosis. Autophagy 12:963–975. https://doi.org/10.1080/15548627.2016.1164359

doi: 10.1080/15548627.2016.1164359 pubmed: 27123694 pmcid: 4922440

Strappazzon F, Di Rita A, Peschiaroli A et al (2019) HUWE1 controls MCL1 stability to unleash AMBRA1-induced mitophagy. Cell Death Differ. https://doi.org/10.1038/s41418-019-0404-8

doi: 10.1038/s41418-019-0404-8 pubmed: 31434979 pmcid: 7206139

Cianfanelli V, Nazio F, Cecconi F (2015) Connecting autophagy: AMBRA1 and its network of regulation. Mol Cell Oncol 2:e970059. https://doi.org/10.4161/23723548.2014.970059

doi: 10.4161/23723548.2014.970059 pubmed: 27308402 pmcid: 4905234

Schoenherr C, Byron A, Griffith B et al (2020) The autophagy protein Ambra1 regulates gene expression by supporting novel transcriptional complexes. J Biol Chem 295:12045–12057. https://doi.org/10.1074/jbc.RA120.012565

doi: 10.1074/jbc.RA120.012565 pubmed: 32616651 pmcid: 7443501

Antonioli M, Albiero F, Nazio F et al (2014) AMBRA1 interplay with cullin E3 Ubiquitin ligases regulates autophagy dynamics. Dev Cell 31:734–746. https://doi.org/10.1016/j.devcel.2014.11.013

doi: 10.1016/j.devcel.2014.11.013 pubmed: 25499913

Di Bartolomeo S, Corazzari M, Nazio F et al (2010) The dynamic interaction of AMBRA1 with the dynein motor complex regulates mammalian autophagy. J Cell Biol 191:155–168. https://doi.org/10.1083/jcb.201002100

doi: 10.1083/jcb.201002100 pubmed: 20921139 pmcid: 2953445

Skobo T, Benato F, Grumati P et al (2014) Zebrafish AMBRA1a and AMBRA1b knockdown impairs skeletal muscle development. PLoS One 9:1–13. https://doi.org/10.1371/journal.pone.0099210

doi: 10.1371/journal.pone.0099210

Maiani E, Milletti G, Nazio F et al (2021) AMBRA1 regulates cyclin D to guard S-phase entry and genomic integrity. Nature 592:799–803. https://doi.org/10.1038/s41586-021-03422-5

doi: 10.1038/s41586-021-03422-5 pubmed: 33854232 pmcid: 8864551

Simoneschi D, Rona G, Zhou N et al (2022) CRL4AMBRA1 is a master regulator of D-type cyclins. Nature 592:789–793. https://doi.org/10.1038/s41586-021-03445-y.CRL4

doi: 10.1038/s41586-021-03445-y.CRL4

Chaikovsky AC, Li C, Jeng EE et al (2021) The AMBRA1 E3 ligase adaptor regulates the stability of cyclin D. Nature 592:794–798. https://doi.org/10.1038/s41586-021-03474-7

doi: 10.1038/s41586-021-03474-7 pubmed: 33854239 pmcid: 8246597

Bayliss R, Fry A, Haq T, Yeoh S (2012) On the molecular mechanisms of mitotic kinase activation. Open Biol 2:120136. https://doi.org/10.1098/rsob.120136

doi: 10.1098/rsob.120136 pubmed: 23226601 pmcid: 3513839

Brown NR, Korolchuk S, Martin MP et al (2015) CDK1 structures reveal conserved and unique features of the essential cell cycle CDK. Nat Commun 6:6769. https://doi.org/10.1038/ncomms7769

doi: 10.1038/ncomms7769 pubmed: 25864384 pmcid: 4413027

Combes G, Alharbi I, Braga LG, Elowe S (2017) Playing polo during mitosis: PLK1 takes the lead. Oncogene 36:4819–4827. https://doi.org/10.1038/onc.2017.113

doi: 10.1038/onc.2017.113 pubmed: 28436952

Lee S-Y, Jang C, Lee K-A (2014) Polo-like kinases (plks), a key regulator of cell cycle and new potential target for cancer therapy. Dev Reprod 18:65–71. https://doi.org/10.12717/DR.2014.18.1.065

doi: 10.12717/DR.2014.18.1.065 pubmed: 25949173 pmcid: 4282265

Yuan K, Huang Y, Yao X (2011) Illumination of mitotic orchestra during cell division: a polo view. Cell Signal 23:1–5. https://doi.org/10.1016/j.cellsig.2010.07.003

doi: 10.1016/j.cellsig.2010.07.003 pubmed: 20633640

Benada J, Burdová K, Lidak T et al (2015) Polo-like kinase 1 inhibits DNA damage response during mitosis. Cell Cycle 14:219–231. https://doi.org/10.4161/15384101.2014.977067

doi: 10.4161/15384101.2014.977067 pubmed: 25607646 pmcid: 4613155

Matthess Y, Raab M, Knecht R et al (2014) Sequential Cdk1 and Plk1 phosphorylation of caspase-8 triggers apoptotic cell death during mitosis. Mol Oncol 8:596–608. https://doi.org/10.1016/j.molonc.2013.12.013

doi: 10.1016/j.molonc.2013.12.013 pubmed: 24484936 pmcid: 5528627

O’Donovan DS, MacFhearraigh S, Whitfield J et al (2013) Sequential Cdk1 and Plk1 phosphorylation of protein tyrosine phosphatase 1B promotes mitotic cell death. Cell Death Dis 4:e468. https://doi.org/10.1038/cddis.2012.208

doi: 10.1038/cddis.2012.208 pubmed: 23348582 pmcid: 3563996

Pintard L, Archambault V (2018) A unified view of spatio-temporal control of mitotic entry: polo kinase as the key. Open Biol 8:180114. https://doi.org/10.1098/rsob.180114

doi: 10.1098/rsob.180114 pubmed: 30135239 pmcid: 6119860

Zhang X, Chen Q, Feng J et al (2009) Sequential phosphorylation of Nedd1 by Cdk1 and Plk1 is required for targeting of the gammaTuRC to the centrosome. J Cell Sci 122:2240–2251. https://doi.org/10.1242/jcs.042747

doi: 10.1242/jcs.042747 pubmed: 19509060

Prosser SL, Pelletier L (2017) Mitotic spindle assembly in animal cells: a fine balancing act. Nat Rev Mol Cell Biol 18:187–201. https://doi.org/10.1038/nrm.2016.162

doi: 10.1038/nrm.2016.162 pubmed: 28174430

Morin X, Bellaïche Y (2011) Mitotic spindle orientation in asymmetric and symmetric cell divisions during animal development. Dev Cell 21:102–119. https://doi.org/10.1016/j.devcel.2011.06.012

doi: 10.1016/j.devcel.2011.06.012 pubmed: 21763612

Radulescu AE, Cleveland DW (2010) NuMA after 30 years: the matrix revisited. Trends Cell Biol 20:214–222. https://doi.org/10.1016/j.tcb.2010.01.003

doi: 10.1016/j.tcb.2010.01.003 pubmed: 20137953 pmcid: 3137513

Du Q, Macara IG (2004) Mammalian Pins is a conformational switch that links NuMA to heterotrimeric G proteins. Cell 119:503–516. https://doi.org/10.1016/j.cell.2004.10.028

doi: 10.1016/j.cell.2004.10.028 pubmed: 15537540

Kotak S, Busso C, Gönczy P (2012) Cortical dynein is critical for proper spindle positioning in human cells. J Cell Biol 199:97–110. https://doi.org/10.1083/jcb.201203166

doi: 10.1083/jcb.201203166 pubmed: 23027904 pmcid: 3461507

Kiyomitsu T, Cheeseman IM (2012) Chromosome- and spindle-pole-derived signals generate an intrinsic code for spindle position and orientation. Nat Cell Biol 14:311–317. https://doi.org/10.1038/ncb2440

doi: 10.1038/ncb2440 pubmed: 22327364 pmcid: 3290711

Seldin L, Muroyama A, Lechler T (2016) NuMA-microtubule interactions are critical for spindle orientation and the morphogenesis of diverse epidermal structures. Elife 5:e12504. https://doi.org/10.7554/eLife.12504.001

doi: 10.7554/eLife.12504.001 pubmed: 26765568 pmcid: 4758947

Pirovano L, Culurgioni S, Carminati M et al (2019) Hexameric NuMA:LGN structures promote multivalent interactions required for planar epithelial divisions. Nat Commun 10:1–14. https://doi.org/10.1038/s41467-019-09999-w

doi: 10.1038/s41467-019-09999-w

Gloerich M, Bianchini JM, Siemers KA et al (2017) Cell division orientation is coupled to cell–cell adhesion by the E-cadherin/LGN complex. Nat Commun 8:1–11. https://doi.org/10.1038/ncomms13996

doi: 10.1038/ncomms13996

Okumura M, Natsume T, Kanemaki MT, Kiyomitsu T (2018) Dynein–dynactin–NuMA clusters generate cortical spindle-pulling forces as a multiarm ensemble. Elife 7:1–24. https://doi.org/10.7554/eLife.36559

doi: 10.7554/eLife.36559

Seldin L, Macara I (2017) Epithelial spindle orientation diversities and uncertainties: recent developments and lingering questions. F1000Research 6:1–9. https://doi.org/10.12688/f1000research.11370.1

doi: 10.12688/f1000research.11370.1

Hueschen CL, Kenny SJ, Xu K, Dumont S (2017) NuMA targets dynein to microtubule minus-ends at mitosis. bioRxiv. https://doi.org/10.1101/148692

doi: 10.1101/148692

Gallini S, Carminati M, De Mattia F et al (2016) NuMA phosphorylation by aurora-a orchestrates spindle orientation. Curr Biol 26:458–469. https://doi.org/10.1016/j.cub.2015.12.051

doi: 10.1016/j.cub.2015.12.051 pubmed: 26832443

Polverino F, Naso FD, Asteriti IA et al (2020) The Aurora-A/TPX2 axis directs spindle orientation in adherent human cells by regulating NuMA and microtubule stability. Curr Biol. https://doi.org/10.1016/j.cub.2020.10.096

doi: 10.1016/j.cub.2020.10.096 pubmed: 33275894

Kotak S, Busso C, Gönczy P (2013) NuMA phosphorylation by CDK1 couples mitotic progression with cortical dynein function. EMBO J 32:2517–2529. https://doi.org/10.1038/emboj.2013.172

doi: 10.1038/emboj.2013.172 pubmed: 23921553 pmcid: 3770949

Sana S, Keshri R, Rajeevan A et al (2018) Plk1 regulates spindle orientation by phosphorylating NuMA in human cells. Life Sci Alliance 1:e201800223. https://doi.org/10.26508/lsa.201800223

doi: 10.26508/lsa.201800223 pubmed: 30456393 pmcid: 6240335

Kõivomägi M, Örd M, Iofik A et al (2013) Multisite phosphorylation networks as signal processors for Cdk1. Nat Struct Mol Biol 20:1415–1424. https://doi.org/10.1038/nsmb.2706.Multisite

doi: 10.1038/nsmb.2706.Multisite pubmed: 24186061

Furuya T, Kim M, Lipinski M et al (2010) Negative regulation of Vps34 by Cdk mediated phosphorylation. Mol Cell 38:500–511. https://doi.org/10.1016/j.molcel.2010.05.009

doi: 10.1016/j.molcel.2010.05.009 pubmed: 20513426 pmcid: 2888511

Elia AEH, Cantley LC, Yaffe MB (2003) Proteomic screen finds pSer/pThr-binding domain localizing Plk1 to mitotic substrates. Science (80–) 299:1228–1231. https://doi.org/10.1126/science.1079079

doi: 10.1126/science.1079079

Strappazzon F, Vietri-Rudan M, Campello S et al (2011) Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J 30:1195–1208. https://doi.org/10.1038/emboj.2011.49

doi: 10.1038/emboj.2011.49 pubmed: 21358617 pmcid: 3094111

Van Humbeeck C, Cornelissen T, Hofkens H et al (2011) Parkin interacts with AMBRA1 to induce mitophagy. J Neurosci 31:10249–10261. https://doi.org/10.1523/JNEUROSCI.1917-11.2011

doi: 10.1523/JNEUROSCI.1917-11.2011 pubmed: 21753002 pmcid: 6623066

Xia P, Wang S, Du Y et al (2013) WASH inhibits autophagy through suppression of Beclin 1 ubiquitination. EMBO J 32:2685–2696. https://doi.org/10.1038/emboj.2013.189

doi: 10.1038/emboj.2013.189 pubmed: 23974797 pmcid: 3801434

Xia P, Wang S, Huang G et al (2014) RNF2 is recruited by WASH to ubiquitinate AMBRA1 leading to downregulation of autophagy. Cell Res 24:943–958. https://doi.org/10.1038/cr.2014.85

doi: 10.1038/cr.2014.85 pubmed: 24980959 pmcid: 4123297

Di Rita A, Peschiaroli A, Acunzo PD et al (2018) HUWE1 E3 ligase promotes PINK1/PARKIN-independent mitophagy by regulating AMBRA1 activation via IKKα. Nat Commun 9:1–18. https://doi.org/10.1038/s41467-018-05722-3

doi: 10.1038/s41467-018-05722-3

Schoenherr C, Byron A, Sandilands E et al (2017) Ambra1 spatially regulates Src activity and Src/FAK-mediated cancer cell invasion via trafficking networks. Elife 6:1–22. https://doi.org/10.7554/eLife.23172

doi: 10.7554/eLife.23172

Cianfanelli V, De Zio D, Di Bartolomeo S et al (2015) AMBRA1 at a glance. J Cell Sci 128:2003–2008. https://doi.org/10.1242/jcs.168153

doi: 10.1242/jcs.168153 pubmed: 26034061

Eskelinen E-L, Prescott AR, Cooper J et al (2002) Inhibition of autophagy in mitotic animal cells. Traffic 3:878–893. https://doi.org/10.1034/j.1600-0854.2002.31204.x

doi: 10.1034/j.1600-0854.2002.31204.x pubmed: 12453151

Li Z, Ji X, Wang D et al (2016) Autophagic flux is highly active in early mitosis and differentially regulated throughout the cell cycle. Oncotarget 12:1–3. https://doi.org/10.18632/oncotarget.9451

doi: 10.18632/oncotarget.9451

Odle RI, Walker SA, Oxley D et al (2019) An mTORC1-to-CDK1 switch maintains autophagy suppression during mitosis. Mol Cell 77:1–13. https://doi.org/10.1016/j.molcel.2019.10.016

doi: 10.1016/j.molcel.2019.10.016

Willson J (2020) Mitosis flips the switch on autophagy control. Nat Rev Mol Cell Biol 21:4–5. https://doi.org/10.1038/s41580-019-0196-1

doi: 10.1038/s41580-019-0196-1 pubmed: 31758162

Li Z, Tian X, Ji X et al (2020) ULK1-ATG13 and their mitotic phospho-regulation by CDK1 connect autophagy to cell cycle. PLoS Biol 18:1–29. https://doi.org/10.1371/journal.pbio.3000288

doi: 10.1371/journal.pbio.3000288

Holdgaard SG, Cianfanelli V, Pupo E et al (2019) Selective autophagy maintains centrosome integrity and accurate mitosis by turnover of centriolar satellites. Nat Commun 10:1–19. https://doi.org/10.1038/s41467-019-12094-9

doi: 10.1038/s41467-019-12094-9

Holdgaard SG, Cianfanelli V, Cecconi F (2019) Cloud hunting: doryphagy, a form of selective macroautophagy that degrades centriolar satellites. Autophagy. https://doi.org/10.1080/15548627.2019.1703356

doi: 10.1080/15548627.2019.1703356 pubmed: 31847687 pmcid: 6984448

Liu L, XIe R, Nguyen S et al (2009) Robust autophagy/mitophagy persists during mitosis Leyuan. Cell Cycle 8:1616–1620. https://doi.org/10.1016/j.pestbp.2011.02.012.Investigations

doi: 10.1016/j.pestbp.2011.02.012.Investigations pubmed: 19411827

Loukil A, Zonca M, Rebouissou C et al (2014) High-resolution live-cell imaging reveals novel cyclin A2 degradation foci involving autophagy. J Cell Sci 127:2145–2150. https://doi.org/10.1242/jcs.139188

doi: 10.1242/jcs.139188 pubmed: 24634511

Linares JF, Amanchy R, Greis K et al (2011) Phosphorylation of p62 by cdk1 controls the timely transit of cells through mitosis and tumor cell proliferation. Mol Cell Biol 31:105–117. https://doi.org/10.1128/MCB.00620-10

doi: 10.1128/MCB.00620-10 pubmed: 20974803

Frémont S, Gérard A, Galloux M et al (2013) Beclin-1 is required for chromosome congression and proper outer kinetochore assembly. EMBO Rep 14:364–372. https://doi.org/10.1038/embor.2013.23

doi: 10.1038/embor.2013.23 pubmed: 23478334 pmcid: 3615652

Maskey D, Yousefi S, Schmid I et al (2013) ATG5 is induced by DNA-damaging agents and promotes mitotic catastrophe independent of autophagy. Nat Commun 4:1–14. https://doi.org/10.1038/ncomms3130

doi: 10.1038/ncomms3130

Woodard GE, Huang N-N, Cho H et al (2010) Ric-8A and Giα recruit LGN, NuMA, and dynein to the cell cortex to help orient the mitotic spindle. Mol Cell Biol 30:3519–3530. https://doi.org/10.1128/mcb.00394-10

doi: 10.1128/mcb.00394-10 pubmed: 20479129 pmcid: 2897540

Couwenbergs C, Labbé JC, Goulding M et al (2007) Heterotrimeric G protein signaling functions with dynein to promote spindle positioning in C. elegans. J Cell Biol 179:15–22. https://doi.org/10.1083/jcb.200707085

doi: 10.1083/jcb.200707085 pubmed: 17908918 pmcid: 2064726

Nguyen-Ngoc T, Afshar K, Gönczy P (2007) Coupling of cortical dynein and Gα proteins mediates spindle positioning in Caenorhabditis elegans. Nat Cell Biol 9:1294–1302. https://doi.org/10.1038/ncb1649

doi: 10.1038/ncb1649 pubmed: 17922003

Keshri R, Rajeevan A, Kotak S (2020) PP2A–B55γ counteracts Cdk1 and regulates proper spindle orientation through the cortical dynein adaptor NuMA. J Cell Sci. https://doi.org/10.1242/jcs.243857

doi: 10.1242/jcs.243857 pubmed: 32591484 pmcid: 7406356

Giráldez S, Galindo-Moreno M, Limón-Mortés MC et al (2017) G1/S phase progression is regulated by PLK1 degradation through the CDK1/βTrCP axis. FASEB J 31:2925–2936. https://doi.org/10.1096/fj.201601108R

doi: 10.1096/fj.201601108R pubmed: 28360195

de Cárcer G, Venkateswaran SV, Salgueiro L et al (2018) Plk1 overexpression induces chromosomal instability and suppresses tumor development. Nat Commun. https://doi.org/10.1038/s41467-018-05429-5

doi: 10.1038/s41467-018-05429-5 pubmed: 30069007 pmcid: 6070485

Quintyne NJ, Reing JE, Hoffelder DR et al (2005) Spindle multipolarity is prevented by centrosomal clustering. Science (80–) 307:127–129. https://doi.org/10.1126/science.1104905

doi: 10.1126/science.1104905

Bergstralh DT, St Johnston D (2014) Spindle orientation: What if it goes wrong? Semin Cell Dev Biol 34:140–145. https://doi.org/10.1016/j.semcdb.2014.06.014

doi: 10.1016/j.semcdb.2014.06.014 pubmed: 24972323 pmcid: 4169663

Bergstralh DT, Dawney NS, St Johnston D (2017) Spindle orientation: a question of complex positioning. Development 144:1137–1145. https://doi.org/10.1242/dev.140764

doi: 10.1242/dev.140764 pubmed: 28351864

Sander JD, Zaback P, Joung JK et al (2007) Zinc Finger Targeter (ZiFiT): an engineered zinc finger/target site design tool. Nucleic Acids Res 35:599–605. https://doi.org/10.1093/nar/gkm349

doi: 10.1093/nar/gkm349

Bae S, Park J, Kim JS (2014) Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases. Bioinformatics 30:1473–1475. https://doi.org/10.1093/bioinformatics/btu048

doi: 10.1093/bioinformatics/btu048 pubmed: 24463181 pmcid: 4016707

Zhou Y, Liu Y, Hussmann D et al (2016) Enhanced genome editing in mammalian cells with a modified dual-fluorescent surrogate system. Cell Mol Life Sci 73:2543–2563. https://doi.org/10.1007/s00018-015-2128-3

doi: 10.1007/s00018-015-2128-3 pubmed: 26755436

Schorl C, Sedivy JM (2007) Analysis of cell cycle phases and progression in cultured mammalian cells. Methods 41:143–150. https://doi.org/10.1016/j.ymeth.2006.07.022

doi: 10.1016/j.ymeth.2006.07.022 pubmed: 17189856 pmcid: 1828876

Wiśniewski JR, Zougman A, Nagaraj N et al (2009) Universal sample preparation method for proteome analysis. Nat Methods 6:359–363. https://doi.org/10.1038/nmeth.1322

doi: 10.1038/nmeth.1322 pubmed: 19377485

Zarei M, Sprenger A, Gretzmeier C, Dengjel J (2013) Rapid combinatorial ERLIC-SCX solid-phase extraction for in-depth phosphoproteome analysis. J Proteome Res 12:5989–5995. https://doi.org/10.1021/pr4007969

doi: 10.1021/pr4007969 pubmed: 24144214

Asteriti IA, Di Cesare E, De Mattia F et al (2014) The Aurora-A inhibitor MLN8237 affects multiple mitotic processes and induces dose-dependent mitotic abnormalities and aneuploidy. Oncotarget 5:6229–6242. https://doi.org/10.18632/oncotarget.2190

doi: 10.18632/oncotarget.2190 pubmed: 25153724 pmcid: 4171625

Liao S, Rajendraprasad G, Wang N et al (2019) Molecular basis of vasohibins-mediated detyrosination and its impact on spindle function and mitosis. Cell Res 29:533–547. https://doi.org/10.1038/s41422-019-0187-y

doi: 10.1038/s41422-019-0187-y pubmed: 31171830 pmcid: 6796878

Lee BH, Schwager F, Meraldi P, Gotta M (2018) p37/UBXN2B regulates spindle orientation by limiting cortical NuMA recruitment via PP1/Repo-Man. J Cell Biol 217:483–593. https://doi.org/10.1083/jcb.201707050

doi: 10.1083/jcb.201707050 pubmed: 29222185 pmcid: 5800812