FLS2 is a CDK-like kinase that directly binds IFT70 and is required for proper ciliary disassembly in Chlamydomonas

English version

Autoři: Qin Zhao ^aff001; Shufen Li ^aff001; Shangjin Shao ^aff001; Zhengmao Wang ^aff001; Junmin Pan ^aff001
Působiště autorů: MOE Key Laboratory of Protein Sciences, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, China ^aff001; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, Shandong Province, China ^aff002
Vyšlo v časopise: FLS2 is a CDK-like kinase that directly binds IFT70 and is required for proper ciliary disassembly in Chlamydomonas. PLoS Genet 16(3): e32767. doi:10.1371/journal.pgen.1008561
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1008561

Souhrn

Intraflagellar transport (IFT) is required for ciliary assembly and maintenance. While disruption of IFT may trigger ciliary disassembly, we show here that IFT mediated transport of a CDK-like kinase ensures proper ciliary disassembly. Mutations in flagellar shortening 2 (FLS2), encoding a CDK-like kinase, lead to retardation of cilia resorption and delay of cell cycle progression. Stimulation for ciliary disassembly induces gradual dephosphorylation of FLS2 accompanied with gradual inactivation. Loss of FLS2 or its kinase activity induces early onset of kinesin13 phosphorylation in cilia. FLS2 is predominantly localized in the cell body, however, it is transported to cilia upon induction of ciliary disassembly. FLS2 directly interacts with IFT70 and loss of this interaction inhibits its ciliary transport, leading to dysregulation of kinesin13 phosphorylation and retardation of ciliary disassembly. Thus, this work demonstrates that IFT plays active roles in controlling proper ciliary disassembly by transporting a protein kinase to cilia to regulate a microtubule depolymerizer.

Klíčová slova:

Cell cycle and cell division – Cilia – Immunoblotting – Immunoprecipitation – In vitro kinase assay – Phosphorylation – Zygotes – Deletion mutagenesis

Zdroje

1. Anvarian Z, Mykytyn K, Mukhopadhyay S, Pedersen LB, Christensen ST. Cellular signalling by primary cilia in development, organ function and disease. Nat Rev Nephrol. 2019;15:199–219. doi: 10.1038/s41581-019-0116-9 30733609

2. Spassky N, Meunier A. The development and functions of multiciliated epithelia. Nat Rev Mol Cell Biol. 2017;18:423–436. doi: 10.1038/nrm.2017.21 28400610

3. Reiter JF, Leroux MR. Genes and molecular pathways underpinning ciliopathies. Nat Rev Mol Cell Biol. 2017;18:533–547. doi: 10.1038/nrm.2017.60 28698599

4. Phua SC, Chiba S, Suzuki M, Su E, Roberson EC, Pusapati GV, et al. Dynamic Remodeling of Membrane Composition Drives Cell Cycle through Primary Cilia Excision. Cell. 2017;168:264–279 e215. doi: 10.1016/j.cell.2016.12.032 28086093

5. Rieder CL, Jensen CG, Jensen LC. The resorption of primary cilia during mitosis in a vertebrate (PtK1) cell line. J Ultrastruct Res. 1979;68:173–185. doi: 10.1016/s0022-5320(79)90152-7 480410

6. Tucker RW, Scher CD, Stiles CD. Centriole deciliation associated with the early response of 3T3 cells to growth factors but not to SV40. Cell. 1979;18:1065–1072. doi: 10.1016/0092-8674(79)90219-8 229969

7. Pugacheva EN, Jablonski SA, Hartman TR, Henske EP, Golemis EA. HEF1-dependent Aurora A activation induces disassembly of the primary cilium. Cell. 2007;129:1351–1363. doi: 10.1016/j.cell.2007.04.035 17604723

8. Liang Y, Meng D, Zhu B, Pan J. Mechanism of ciliary disassembly. Cell Mol Life Sci. 2016;73:1787–1802. doi: 10.1007/s00018-016-2148-7 26869233

9. Hsu KS, Chuang JZ, Sung CH. The Biology of Ciliary Dynamics. Cold Spring Harb Perspect Biol. 2017;9.

10. Korobeynikov V, Deneka AY, Golemis EA. Mechanisms for nonmitotic activation of Aurora-A at cilia. Biochem Soc Trans. 2017;45:37–49. doi: 10.1042/BST20160142 28202658

11. Quarmby LM. Cellular deflagellation. Int Rev Cytol. 2004;233:47–91. doi: 10.1016/S0074-7696(04)33002-0 15037362

12. Bloodgood RA. Resorption of organelles containing microtubules. Cytobios. 1974;9:142–161. 4603233

13. Hu Z, Liang Y, He W, Pan J. Cilia disassembly with two distinct phases of regulation. Cell Rep. 2015;10:1803–1810. doi: 10.1016/j.celrep.2015.02.044 25801021

14. Cavalier-Smith T. Basal body and flagellar development during the vegetative cell cycle and the sexual cycle of Chlamydomonas reinhardii. J Cell Sci. 1974;16:529–556. 4615103

15. Mirvis M, Siemers KA, Nelson WJ, Stearns TP. Primary cilium loss in mammalian cells occurs predominantly by whole-cilium shedding. PLoS Biol. 2019;17:e3000381. doi: 10.1371/journal.pbio.3000381 31314751

16. Izawa I, Goto H, Kasahara K, Inagaki M. Current topics of functional links between primary cilia and cell cycle. Cilia. 2015;4:12. doi: 10.1186/s13630-015-0021-1 26719793

17. Kim S, Zaghloul NA, Bubenshchikova E, Oh EC, Rankin S, Katsanis N, et al. Nde1-mediated inhibition of ciliogenesis affects cell cycle re-entry. Nat Cell Biol. 2011;13:351–360. doi: 10.1038/ncb2183 21394081

18. Li A, Saito M, Chuang JZ, Tseng YY, Dedesma C, Tomizawa K, et al. Ciliary transition zone activation of phosphorylated Tctex-1 controls ciliary resorption, S-phase entry and fate of neural progenitors. Nat Cell Biol. 2011;13:402–411. doi: 10.1038/ncb2218 21394082

19. Jackson PK. Do cilia put brakes on the cell cycle? Nat Cell Biol. 2011;13:340–342. doi: 10.1038/ncb0411-340 21460803

20. Zhang W, Yang SL, Yang M, Herrlinger S, Shao Q, Collar JL, et al. Modeling microcephaly with cerebral organoids reveals a WDR62-CEP170-KIF2A pathway promoting cilium disassembly in neural progenitors. Nat Commun. 2019;10:2612. doi: 10.1038/s41467-019-10497-2 31197141

21. Gabriel E, Wason A, Ramani A, Gooi LM, Keller P, Pozniakovsky A, et al. CPAP promotes timely cilium disassembly to maintain neural progenitor pool. EMBO J. 2016;35:803–819. doi: 10.15252/embj.201593679 26929011

22. Eguether T, Hahne M. Mixed signals from the cell's antennae: primary cilia in cancer. EMBO Rep. 2018;19.

23. Pan J, Wang Q, Snell WJ. An aurora kinase is essential for flagellar disassembly in Chlamydomonas. Dev Cell. 2004;6:445–451. doi: 10.1016/s1534-5807(04)00064-4 15030766

24. Miyamoto T, Hosoba K, Ochiai H, Royba E, Izumi H, Sakuma T, et al. The Microtubule-Depolymerizing Activity of a Mitotic Kinesin Protein KIF2A Drives Primary Cilia Disassembly Coupled with Cell Proliferation. Cell Rep. 2015;10:664–673. doi: 10.1016/j.celrep.2015.01.003 25660017

25. Piao T, Luo M, Wang L, Guo Y, Li D, Li P, et al. A microtubule depolymerizing kinesin functions during both flagellar disassembly and flagellar assembly in Chlamydomonas. Proc Natl Acad Sci U S A. 2009;106:4713–4718. doi: 10.1073/pnas.0808671106 19264963

26. Kim S, Lee K, Choi JH, Ringstad N, Dynlacht BD. Nek2 activation of Kif24 ensures cilium disassembly during the cell cycle. Nat Commun. 2015;6:8087. doi: 10.1038/ncomms9087 26290419

27. Scholey JM. Intraflagellar transport. Annu Rev Cell Dev Biol. 2003;19:423–443. doi: 10.1146/annurev.cellbio.19.111401.091318 14570576

28. Rosenbaum JL, Witman GB. Intraflagellar transport. Nat Rev Mol Cell Biol. 2002;3:813–825. doi: 10.1038/nrm952 12415299

29. Lux FG 3rd, Dutcher SK. Genetic interactions at the FLA10 locus: suppressors and synthetic phenotypes that affect the cell cycle and flagellar function in Chlamydomonas reinhardtii. Genetics. 1991;128:549–561. 1874415

30. Engelke MF, Waas B, Kearns SE, Suber A, Boss A, Allen BL, et al. Acute Inhibition of Heterotrimeric Kinesin-2 Function Reveals Mechanisms of Intraflagellar Transport in Mammalian Cilia. Curr Biol. 2019;29:1137–1148 e1134. doi: 10.1016/j.cub.2019.02.043 30905605

31. Mueller J, Perrone CA, Bower R, Cole DG, Porter ME. The FLA3 KAP subunit is required for localization of kinesin-2 to the site of flagellar assembly and processive anterograde intraflagellar transport. Mol Biol Cell. 2005;16:1341–1354. doi: 10.1091/mbc.E04-10-0931 15616187

32. Pan J, Snell WJ. Chlamydomonas shortens its flagella by activating axonemal disassembly, stimulating IFT particle trafficking, and blocking anterograde cargo loading. Dev Cell. 2005;9:431–438. doi: 10.1016/j.devcel.2005.07.010 16139231

33. Wang L, Gu L, Meng D, Wu Q, Deng H, Pan J. Comparative Proteomics Reveals Timely Transport into Cilia of Regulators or Effectors as a Mechanism Underlying Ciliary Disassembly. J Proteome Res. 2017;16:2410–2418. doi: 10.1021/acs.jproteome.6b01048 28534617

34. Lefebvre PA, Nordstrom SA, Moulder JE, Rosenbaum JL. Flagellar elongation and shortening in Chlamydomonas. IV. Effects of flagellar detachment, regeneration, and resorption on the induction of flagellar protein synthesis. J Cell Biol. 1978;78:8–27. doi: 10.1083/jcb.78.1.8 149796

35. Endicott JA, Noble ME. Structural characterization of the cyclin-dependent protein kinase family. Biochem Soc Trans. 2013;41:1008–1016. doi: 10.1042/BST20130097 23863171

36. Yee KW, Moore SJ, Midmer M, Zanke BW, Tong F, Hedley D, et al. NKIAMRE, a novel conserved CDC2-related kinase with features of both mitogen-activated protein kinases and cyclin-dependent kinases. Biochem Biophys Res Commun. 2003;308:784–792. doi: 10.1016/s0006-291x(03)01475-x 12927787

37. Horinouchi T, Terada K, Higashi T, Miwa S. Using Phos-Tag in Western Blotting Analysis to Evaluate Protein Phosphorylation. Methods Mol Biol. 2016;1397:267–277. doi: 10.1007/978-1-4939-3353-2_18 26676139

38. Kinoshita E, Kinoshita-Kikuta E, Takiyama K, Koike T. Phosphate-binding tag, a new tool to visualize phosphorylated proteins. Mol Cell Proteomics. 2006;5:749–757. doi: 10.1074/mcp.T500024-MCP200 16340016

39. Cao M, Meng D, Wang L, Bei S, Snell WJ, Pan J. Activation loop phosphorylation of a protein kinase is a molecular marker of organelle size that dynamically reports flagellar length. Proc Natl Acad Sci U S A. 2013;110:12337–12342. doi: 10.1073/pnas.1302364110 23836633

40. Qin H, Diener DR, Geimer S, Cole DG, Rosenbaum JL. Intraflagellar transport (IFT) cargo: IFT transports flagellar precursors to the tip and turnover products to the cell body. J Cell Biol. 2004;164:255–266. doi: 10.1083/jcb.200308132 14718520

41. Wingfield JL, Lechtreck KF, Lorentzen E. Trafficking of ciliary membrane proteins by the intraflagellar transport/BBSome machinery. Essays Biochem. 2018;62:753–763. doi: 10.1042/EBC20180030 30287585

42. Lechtreck KF. IFT-Cargo Interactions and Protein Transport in Cilia. Trends Biochem Sci. 2015;40:765–778. doi: 10.1016/j.tibs.2015.09.003 26498262

43. Kee HL, Dishinger JF, Blasius TL, Liu CJ, Margolis B, Verhey KJ. A size-exclusion permeability barrier and nucleoporins characterize a ciliary pore complex that regulates transport into cilia. Nat Cell Biol. 2012;14:431–437. doi: 10.1038/ncb2450 22388888

44. Walther Z, Vashishtha M, Hall JL. The Chlamydomonas FLA10 gene encodes a novel kinesin-homologous protein. J Cell Biol. 1994;126:175–188. doi: 10.1083/jcb.126.1.175 8027176

45. Kozminski KG, Beech PL, Rosenbaum JL. The Chlamydomonas kinesin-like protein FLA10 is involved in motility associated with the flagellar membrane. J Cell Biol. 1995;131:1517–1527. doi: 10.1083/jcb.131.6.1517 8522608

46. Taschner M, Kotsis F, Braeuer P, Kuehn EW, Lorentzen E. Crystal structures of IFT70/52 and IFT52/46 provide insight into intraflagellar transport B core complex assembly. J Cell Biol. 2014;207:269–282. doi: 10.1083/jcb.201408002 25349261

47. Takei R, Katoh Y, Nakayama K. Robust interaction of IFT70 with IFT52-IFT88 in the IFT-B complex is required for ciliogenesis. Biol Open. 2018;7.

48. Taschner M, Lorentzen E. The Intraflagellar Transport Machinery. Cold Spring Harb Perspect Biol. 2016;8.

49. Marshall WF, Rosenbaum JL. Intraflagellar transport balances continuous turnover of outer doublet microtubules: implications for flagellar length control. J Cell Biol. 2001;155:405–414. doi: 10.1083/jcb.200106141 11684707

50. Wang L, Piao T, Cao M, Qin T, Huang L, Deng H, et al. Flagellar regeneration requires cytoplasmic microtubule depolymerization and kinesin-13. J Cell Sci. 2013;126:1531–1540. doi: 10.1242/jcs.124255 23418346

51. Dubos A, Pannetier S, Hanauer A. Inactivation of the CDKL3 gene at 5q31.1 by a balanced t(X;5) translocation associated with nonspecific mild mental retardation. Am J Med Genet A. 2008;146A:1267–1279. doi: 10.1002/ajmg.a.32274 18412109

52. Kilstrup-Nielsen C, Rusconi L, La Montanara P, Ciceri D, Bergo A, Bedogni F, et al. What we know and would like to know about CDKL5 and its involvement in epileptic encephalopathy. Neural Plast. 2012;2012:728267. doi: 10.1155/2012/728267 22779007

53. Zhu B, Zhu X, Wang L, Liang Y, Feng Q, Pan J. Functional exploration of the IFT-A complex in intraflagellar transport and ciliogenesis. PLoS Genet. 2017;13:e1006627. doi: 10.1371/journal.pgen.1006627 28207750

54. Bisova K, Krylov DM, Umen JG. Genome-wide annotation and expression profiling of cell cycle regulatory genes in Chlamydomonas reinhardtii. Plant Physiol. 2005;137:475–491. doi: 10.1104/pp.104.054155 15710686

55. Zhu X, Liang Y, Gao F, Pan J. IFT54 regulates IFT20 stability but is not essential for tubulin transport during ciliogenesis. Cell Mol Life Sci. 2017;74:3425–3437. doi: 10.1007/s00018-017-2525-x 28417161

56. Meng D, Cao M, Oda T, Pan J. The conserved ciliary protein Bug22 controls planar beating of Chlamydomonas flagella. J Cell Sci. 2014;127:281–287. doi: 10.1242/jcs.140723 24259666

57. Berthold P, Schmitt R, Mages W. An engineered Streptomyces hygroscopicus aph 7" gene mediates dominant resistance against hygromycin B in Chlamydomonas reinhardtii. Protist. 2002;153:401–412. doi: 10.1078/14344610260450136 12627869

58. Liang Y, Pan J. Regulation of flagellar biogenesis by a calcium dependent protein kinase in Chlamydomonas reinhardtii. PLoS One. 2013;8:e69902. doi: 10.1371/journal.pone.0069902 23936117

59. Molnar A, Bassett A, Thuenemann E, Schwach F, Karkare S, Ossowski S, et al. Highly specific gene silencing by artificial microRNAs in the unicellular alga Chlamydomonas reinhardtii. Plant J. 2009;58:165–174. doi: 10.1111/j.1365-313X.2008.03767.x 19054357

60. Lv B, Wan L, Taschner M, Cheng X, Lorentzen E, Huang K. Intraflagellar transport protein IFT52 recruits IFT46 to the basal body and flagella. J Cell Sci. 2017;130:1662–1674. doi: 10.1242/jcs.200758 28302912

61. Bloodgood RA, Salomonsky NL. The transmembrane signaling pathway involved in directed movements of Chlamydomonas flagellar membrane glycoproteins involves the dephosphorylation of a 60-kD phosphoprotein that binds to the major flagellar membrane glycoprotein. J Cell Biol. 1994;127:803–811. doi: 10.1083/jcb.127.3.803 7962061

62. Allen JJ, Li M, Brinkworth CS, Paulson JL, Wang D, Hubner A, et al. A semisynthetic epitope for kinase substrates. Nat Methods. 2007;4:511–516. doi: 10.1038/nmeth1048 17486086

Článek Bayesian network analysis incorporating genetic anchors complements conventional Mendelian randomization approaches for exploratory analysis of causal relationships in complex data

Článek Disentangling group specific QTL allele effects from genetic background epistasis using admixed individuals in GWAS: An application to maize flowering

Článek The Lid/KDM5 histone demethylase complex activates a critical effector of the oocyte-to-zygote transition

Článek The emerging role of transcriptional regulation in the oocyte-to-zygote transition

Článek Murine cytomegalovirus infection exacerbates complex IV deficiency in a model of mitochondrial disease

Článek The SNAP hypothesis: Chromosomal rearrangements could emerge from positive Selection during Niche Adaptation

Článek Inhibition of the oligosaccharyl transferase in Caenorhabditis elegans that compromises ER proteostasis suppresses p38-dependent protection against pathogenic bacteria

Článek Histone H4 dosage modulates DNA damage response in the pathogenic yeast Candida glabrata via homologous recombination pathway

Článek Correction: Metagenomic sequencing suggests a diversity of RNA interference-like responses to viruses across multicellular eukaryotes

Článek Drosophila insulin-like peptide 2 mediates dietary regulation of sleep intensity

Článek The alarmones (p)ppGpp are part of the heat shock response of Bacillus subtilis

Článek RNA Polymerase II CTD phosphatase Rtr1 fine-tunes transcription termination