Centrosome-derived microtubule radial array, PCM-1 protein, and primary cilia formation
In animal cells, the centrosome nucleates and anchors microtubules (MT), forming their radial array. During interphase centrosome-derived MT, aster can either team up with other MT network or function in an autonomous manner. What is the function of the centrosome-derived MT aster? We suggested that it might play an important role in the formation of the primary cilium, the organelle obligatorily associated with the centrosome. PCM-1 (PeriCentriolar Matrix 1) protein, which participates in the organization of the primary cilium, is a part of pericentiolar satellites. They are transported to the centrosome along MTs by the motor protein dynein in a complex with its cofactor dynactin. Previously, we showed that SLK/LOSK phosphorylated the p150Glued subunit of dynactin, thus promoting its centrosomal targeting followed by its participation in the retention of microtubules. Here, we found that under the repression of SLK/LOSK activity, the PCM-1 protein lost its pericentrosomal localization and was being dispersed throughout the cytoplasm. Despite that the alanine and glutamine mutants of p150Glued had opposite effects on PCM-1 localization, they associated with PCM-1 to the same extent. The occurrence of primary cilia also significantly decreased when SLK/LOSK was repressed. These defects also correlated with a disturbance of the long-range transport in cells, whereas dynein-depending motility was intact. Treatment with the GSK-3β kinase inhibitor also resulted in the loss of the centrosome-derived MT aster, dispersion of PCM-1 over the cytoplasm, and reduction of primary cilia occurrence. Thus, kinases involved in the centrosome-derived MT aster regulation can indirectly control the formation of primary cilia in cells.
KeywordsProtein kinase Dynactin GSK-3β SLK LOSK Dynein-driven transport PCM-1 Primary cilium
The authors are very grateful to Prof. Andrea Merdes (Université Paul Sabatier/CNRS, Toulouse) for the great anti-PCM-1 antibody. We also thank Dr. Hans-Peter Hauri (University of Basel) for DNA construct, Dr. Ilya Brodsky (Moscow State University) for DNA constructs and fruitful discussions, and Ekaterina Ryabkova for technical assistance.
The project was financially supported by the President of Russain Federation grant for young PhD researchers (MK-8703.2010.4) and grants from the Russian Basic Research Foundation (08-04-01697-a, 11-04-01022-a, and 18-04-00742-a).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Cantagrel V, Silhavy JL, Bielas SL, Swistun D, Marsh SE, Bertrand JY, Audollent S, Attié-Bitach T, Holden KR, Dobyns WB, Traver D, Al-Gazali L, Ali BR, Lindner TH, Caspary T, Otto EA, Hildebrandt F, Glass IA, Logan CV, Johnson CA, Bennett C, Brancati F, International Joubert Syndrome Related Disorders Study Group, Valente EM, Woods CG, Gleeson JG (2008) Mutations in the cilia gene ARL13B lead to the classical form of Joubert syndrome. Am J Hum Genet 83:170–179. https://doi.org/10.1016/j.ajhg.2008.06.023 CrossRefPubMedPubMedCentralGoogle Scholar
- Fargier G, Favard C, Parmeggiani A, Sahuquet A, Mérezègue F, Morel A, Denis M, Molinari N, Mangeat PH, Coopman PJ, Montcourrier P (2013) Centrosomal targeting of Syk kinase is controlled by its catalytic activity and depends on MTs and the dynein motor. FASEB J 27:109–122. https://doi.org/10.1096/fj.11-202465 CrossRefPubMedGoogle Scholar
- Gao FJ, Hebbar S, Gao XA, Alexander M, Pandey JP, Walla MD, Cotham WE, King SJ, Smith DS (2015) GSK-3β phosphorylation of cytoplasmic dynein reduces Ndel1 binding to intermediate chains and alters dynein motility. Traffic 16:941–961. https://doi.org/10.1111/tra.12304 CrossRefPubMedPubMedCentralGoogle Scholar
- Guilluy C, Rolli-Derkinderen M, Loufrani L, Bourge A, Henrion D, Sabourin L, Loirand G, Pacaud P (2008) Ste20-related kinase SLK phosphorylates Ser188 of RhoA to induce vasodilation in response to angiotensin II type 2 receptor activation. Circ Res 102:1265–1274. https://doi.org/10.1161/CIRCRESAHA.107.164764 CrossRefPubMedGoogle Scholar
- Gupta GD, Coyaud É, Gonçalves J, Mojarad BA, Liu Y, Wu Q, Gheiratmand L, Comartin D, Tkach JM, Cheung SW, Bashkurov M, Hasegan M, Knight JD, Lin ZY, Schueler M, Hildebrandt F, Moffat J, Gingras AC, Raught B, Pelletier L (2015) A dynamic protein interaction landscape of the human centrosome-cilium interface. Cell 163:1484–1499. https://doi.org/10.1016/j.cell.2015.10.065 CrossRefPubMedPubMedCentralGoogle Scholar
- Johnson TM, Antrobus R, Johnson LN (2008) Plk1 activation by Ste20-like kinase (Slk) phosphorylation and polo-box phosphopeptide binding assayed with the substrate translationally controlled tumor protein (TCTP). Biochemistry 47:3688–3696. https://doi.org/10.1021/bi702134c CrossRefPubMedGoogle Scholar
- Lopes CA, Prosser SL, Romio L, Hirst RA, O’Callaghan C, Woolf AS, Fry AM (2011) Centriolar satellites are assembly points for proteins implicated in human ciliopathies, including oral-facial-digital syndrome 1. J Cell Sci 124:600–612. https://doi.org/10.1242/jcs.077156 CrossRefPubMedPubMedCentralGoogle Scholar
- Mariappan A, Soni K, Schorpp K, Zhao F, Minakar A, Zheng X, Mandad S, Macheleidt I, Ramani A, Kubelka T, Dawidowski M, Golfmann K, Wason A, Yang C, Simons J, Schmalz HG, Hyman AA, Aneja R, Ullrich R, Urlaub H, Odenthal M, Büttner R, Li H, Sattler M, Hadian K, Gopalakrishnan J (2019) Inhibition of CPAP-tubulin interaction prevents proliferation of centrosome-amplified cancer cells. EMBO J 38:e99876. https://doi.org/10.15252/embj.201899876 CrossRefPubMedGoogle Scholar
- Olsen JV, Vermeulen M, Santamaria A, Kumar C, Miller ML, Jensen LJ, Gnad F, Cox J, Jensen TS, Nigg EA, Brunak S, Mann M (2010) Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during mitosis. Sci Signal 3(104):ra3. https://doi.org/10.1126/scisignal.2000475 CrossRefPubMedPubMedCentralGoogle Scholar
- Staples CJ, Myers KN, Beveridge RD, Patil AA, Howard AE, Barone G, Lee AJ, Swanton C, Howell M, Maslen S, Skehel JM, Boulton SJ, Collis SJ (2014) Ccdc13 is a novel human centriolar satellite protein required for ciliogenesis and genome stability. J Cell Sci 127:2910–2919. https://doi.org/10.1242/jcs.147785 CrossRefPubMedGoogle Scholar
- Tripathy SK, Weil SJ, Chen C, Anand P, Vallee RB, Gross SP (2014) Autoregulatory mechanism for dynactin control of processive and diffusive dynein transport. Nat Cell Biol 16:1192–1201. https://doi.org/10.1038/ncb3063
- Wheway G, Nazlamova L, Hancock JT (2018) Signaling through the primary cilium. Front Cell Dev Biol 6(8). https://doi.org/10.3389/fcell.2018.00008