The Caenorhabditis elegans one-cell embryo polarizes in response to a cue from the paternally donated centrosome and asymmetrically segregates cell fate determinants that direct the developmental program of the worm. We have found that genes encoding putative deubiquitylating enzymes (DUBs) are required for polarization of one-cell embryos. Maternal loss of the proteins MATH-33 and USP-47 leads to variable inability to correctly establish and maintain asymmetry as defined by posterior and anterior polarity proteins PAR-2 and PAR-3. The first observable defect is variable positioning of the centrosome with respect to the cell cortex and the male pronucleus. The severity of the polarity defects correlates with distance of the centrosome from the cortex. Furthermore, polarity defects can be bypassed by mutations that bring the centrosome in close proximity to the cortex. In addition we find that polarity and centrosome positioning defects can be suppressed by compromising protein turnover. We propose that the DUB activity of MATH-33 and USP-47 stabilizes one or more proteins required for association of the centrosome with the cortex. Because these DUBs are homologous to two members of a group of DUBs that act in fission yeast polarity, we tested additional members of that family and found that another C. elegans DUB gene, usp-46, also contributes to polarity. Our finding that deubiquitylating enzymes required for polarity in Schizosaccharomyces pombe are also required in C. elegans raises the possibility that these DUBs act through an evolutionarily conserved mechanism to control cell polarity.
Vyšlo v časopise: Deubiquitylation Machinery Is Required for Embryonic Polarity in. PLoS Genet 8(11): e32767. doi:10.1371/journal.pgen.1003092 Kategorie: Research Article prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003092
1. KemphuesKJ, PriessJR, MortonDG, ChengNS (1988) Identification of genes required for cytoplasmic localization in early C. elegans embryos. Cell 52 : 311–320.
2. GoldsteinB, MacaraIG (2007) The PAR proteins: fundamental players in animal cell polarization. Developmental cell 13 : 609–622.
3. KemphuesK (2000) PARsing Embryonic Polarity. Cell 101 : 345–348.
4. GuoS, KemphuesKJ (1995) par-1, a gene required for establishing polarity in C. elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed. Cell 81 : 611–620.
5. BoydL, GuoS, LevitanD, StinchcombDT, KemphuesKJ (1996) PAR-2 is asymmetrically distributed and promotes association of P granules and PAR-1 with the cortex in C. elegans embryos. Development 122 : 3075–3084.
6. BeattyA, MortonD, KemphuesK (2010) The C. elegans homolog of Drosophila Lethal giant larvae functions redundantly with PAR-2 to maintain polarity in the early embryo. Development 137 : 3995–4004.
7. HoegeC, ConstantinescuA-T, SchwagerA, GoehringNW, KumarP, et al. (2010) LGL can partition the cortex of one-cell Caenorhabditis elegans embryos into two domains. Current Biology 20 : 1296–1303.
8. St JohnstonD, AhringerJ (2010) Cell polarity in eggs and epithelia: parallels and diversity. Cell 141 : 757–774.
9. CowanCR, HymanAA (2007) Acto-myosin reorganization and PAR polarity in C. elegans. Development 134 : 1035–1043.
10. CuencaAA, SchetterA, AcetoD, KemphuesK, SeydouxG (2003) Polarization of the C. elegans zygote proceeds via distinct establishment and maintenance phases. Development 130 : 1255–1265.
11. CowanCR, HymanAA (2004) Centrosomes direct cell polarity independently of microtubule assembly in C. elegans embryos. Nature 431 : 92–96.
12. MunroE, NanceJ, PriessJR (2004) Cortical flows powered by asymmetrical contraction transport PAR proteins to establish and maintain anterior-posterior polarity in the early C. elegans embryo. Developmental cell 7 : 413–424.
13. TsaiM-C, AhringerJ (2007) Microtubules are involved in anterior-posterior axis formation in C. elegans embryos. The Journal of cell biology 179 : 397–402.
14. MotegiF, SugimotoA (2006) Sequential functioning of the ECT-2 RhoGEF, RHO-1 and CDC-42 establishes cell polarity in Caenorhabditis elegans embryos. Nature cell biology 8 : 978–985.
15. SchoneggS, HymanAA (2006) CDC-42 and RHO-1 coordinate acto-myosin contractility and PAR protein localization during polarity establishment in C. elegans embryos. Development 133 : 3507–3516.
16. JenkinsN, SaamJR, MangoSE (2006) CYK-4/GAP provides a localized cue to initiate anteroposterior polarity upon fertilization. Science 313 : 1298–1301.
17. ZoniesS, MotegiF, HaoY, SeydouxG (2010) Symmetry breaking and polarization of the C. elegans zygote by the polarity protein PAR-2. Development (Cambridge, England) 137 : 1669–1677.
18. MotegiF, ZoniesS, HaoY, CuencaA, GriffinE, et al. (2011) Microtubules induce self-organization of polarized PAR domains in Caenorhabditis elegans zygotes. Nature cell biology 13 : 1361–1367.
19. CheeksRJ, CanmanJC, GabrielWN, MeyerN, StromeS, et al. (2004) C. elegans PAR Proteins Function by Mobilizing and Stabilizing Asymmetrically Localized Protein Complexes. Current Biology 14 : 851–862.
20. SchumacherJM, AshcroftN, DonovanPJ, Goldena (1998) A highly conserved centrosomal kinase, AIR-1, is required for accurate cell cycle progression and segregation of developmental factors in Caenorhabditis elegans embryos. Development (Cambridge, England) 125 : 4391–4402.
21. O'ConnellKF, MaxwellKN, WhiteJG (2000) The spd-2 gene is required for polarization of the anteroposterior axis and formation of the sperm asters in the Caenorhabditis elegans zygote. Developmental biology 222 : 55–70.
22. HamillDR, SeversonAF, CarterJC, BowermanB (2002) Centrosome maturation and mitotic spindle assembly in C. elegans require SPD-5, a protein with multiple coiled-coil domains. Developmental cell 3 : 673–684.
23. LyczakR, ZweierL, GroupT, MurrowMA, SnyderC, et al. (2006) The puromycin-sensitive aminopeptidase PAM-1 is required for meiotic exit and anteroposterior polarity in the one-cell Caenorhabditis elegans embryo. Development 133 : 4281–4292.
24. FortinSM, MarshallSL, JaegerEC, GreenePE, BradyLK, et al. (2010) The PAM-1 aminopeptidase regulates centrosome positioning to ensure anterior-posterior axis specification in one-cell C. elegans embryos. Developmental biology 344 : 992–1000.
25. HaoY, BoydL, SeydouxG (2006) Stabilization of cell polarity by the C. elegans RING protein PAR-2. Developmental Cell 199–208.
26. MortonDG, ShakesDC, NugentS, DichosoD, WangW, et al. (2002) The Caenorhabditis elegans par-5 gene encodes a 14-3-3 protein required for cellular asymmetry in the early embryo. Developmental biology 241 : 47–58.
27. KipreosET (2005) Ubiquitin-mediated pathways in C. elegans. Worm Book: the online review of C elegans biology 1–24.
28. HershkoA, CiechanoverA (1992) The ubiquitin system for protein degradation. Annual review of biochemistry 61 : 761–807.
29. RaiborgC, StenmarkH (2009) The ESCRT machinery in endosomal sorting of ubiquitylated membrane proteins. Nature 458 : 445–452.
30. VentiiKH, WilkinsonKD (2008) Protein partners of deubiquitinating enzymes. The Biochemical journal 414 : 161–175.
31. MortonDG, HooseWA, KemphuesKJ (2012) A genome-wide RNAi screen for enhancers of Par mutants reveals new contributors to early embryonic polarity in Caenorhabditis elegans. Genetics 192 in press.
32. MiwaJ, SchierenbergE, MiwaS, von EhrensteinG (1980) Genetics and mode of expression of temperature-sensitive mutations arresting embryonic development in Caenorhabditis elegans. Developmental biology 76 : 160–174.
33. WoodWB, HechtR, CarrS, VandersliceR, WolfN, et al. (1980) Parental effects and phenotypic characterization of mutations that affect early development in Caenorhabditis elegans. Developmental biology 74 : 446–469.
34. KemphuesKJ, WolfN, WoodWB, HirshD (1986) Two loci required for cytoplasmic organization in early embryos of Caenorhabditis elegans. Developmental biology 113 : 449–460.
35. NakamuraK, KimS, IshidateT, BeiY, PangK, et al. (2005) Wnt signaling drives WRM-1/beta-catenin asymmetries in early C. elegans embryos. Genes & development 19 : 1749–1754.
36. WalstonT, GuoC, ProencaR, WuM, HermanM, et al. (2006) mig-5/Dsh controls cell fate determination and cell migration in C. elegans. Developmental biology 298 : 485–497 Available:http://www.ncbi.nlm.nih.gov/pubmed/16899238.
37. LevitanDJ, BoydL, MelloCC, KemphuesKJ, StinchcombDT (1994) par-2, a gene required for blastomere asymmetry in Caenorhabditis elegans, encodes zinc-finger and ATP-binding motifs. Proceedings of the National Academy of Sciences of the United States of America 91 : 6108–6112.
38. NanceJ, MunroEM, PriessJR (2003) C. elegans PAR-3 and PAR-6 are required for apicobasal asymmetries associated with cell adhesion and gastrulation. Development 130 : 5339–5350.
39. MayorT, LipfordJR, GraumannJ, SmithGT, DeshaiesRJ (2005) Analysis of polyubiquitin conjugates reveals that the Rpn10 substrate receptor contributes to the turnover of multiple proteasome targets. Molecular & cellular proteomics: MCP 4 : 741–751.
40. DavyA, BelloP, Thierry-MiegN, VaglioP, HittiJ, et al. (2001) A protein-protein interaction map of the Caenorhabditis elegans 26S proteasome. EMBO reports 2 : 821–828.
41. ShimadaM, KanematsuK, TanakaK, YokosawaH, KawaharaH (2006) Proteasomal Ubiquitin Receptor RPN-10 Controls Sex Determination in Caenorhabditis elegans. Molecular Biology of the Cell 17 : 5356–5371.
42. LabbéJ-C, PacqueletA, MartyT, GottaM (2006) A genomewide screen for suppressors of par-2 uncovers potential regulators of PAR protein-dependent cell polarity in Caenorhabditis elegans. Genetics 174 : 285–295.
43. KourantiI, McLeanJR, FeoktistovaA, LiangP, JohnsonAE, et al. (2010) A global census of fission yeast deubiquitinating enzyme localization and interaction networks reveals distinct compartmentalization profiles and overlapping functions in endocytosis and polarity. PLoS Biol 8: e1000471 doi:10.1371/journal.pbio.1000471
44. PacqueletA, ZaninE, AshionoC, GottaM (2008) PAR-6 levels are regulated by NOS-3 in a CUL-2 dependent manner in Caenorhabditiselegans. Developmental biology 319 : 267–272.
45. HyenneV, DesrosiersM, LabbéJ-C (2008) C. elegans Brat homologs regulate PAR protein-dependent polarity and asymmetric cell division. Developmental biology 321 : 368–378.
46. SugiyamaY, NishimuraA, OhnoS (2008) Symmetrically dividing cell specific division axes alteration observed in proteasome depleted C. elegans embryo. Mechanisms of development 125 : 743–755.
47. BienkowskaD, CowanCR (2012) Centrosomes Can Initiate a Polarity Axis from Any Position within One-Cell C. elegans Embryos. Current Biology 1–7.
48. ReinschS, GönczyP (1998) Mechanisms of nuclear positioning. Journal of cell science 111 : 2283–2295.
49. CockellMM, BaumerK, GönczyP (2004) lis-1 is required for dynein-dependent cell division processes in C. elegans embryos. Journal of Cell Science 117 : 4571–4582.
50. NockerS van, SadisS, RubinD, GlickmanM, FuH, et al. (1996) The multiubiquitin-chain-binding protein Mcb1 is a component of the 26S proteasome in Saccharomyces cerevisiae and plays a nonessential, substrate-specific role in protein turnover. Molecular and Cellular Biology 16 : 6020–6028.
51. SiegristSE, DoeCQ (2007) Microtubule-induced cortical cell polarity. Genes & development 21 : 483–496.
52. MartinSG (2009) Microtubule-dependent cell morphogenesis in the fission yeast. Trends in cell biology 19 : 447–454.
53. KahnNW, ReaSL, MoyleS, KellA, JohnsonTE (2008) Proteasomal dysfunction activates the transcription factor SKN-1 and produces a selective oxidative-stress response in Caenorhabditis elegans. The Biochemical journal 409 : 205–213.
54. RocheleauCE, CullisonK, HuangK, BernsteinY, SpilkerAC, et al. (2008) The Caenorhabditis elegans ekl (enhancer of ksr-1 lethality) genes include putative components of a germline small RNA pathway. Genetics 178 : 1431–1443.
55. ZahreddineH, ZhangH, DiogonM, NagamatsuY, LabouesseM (2010) CRT-1/calreticulin and the E3 ligase EEL-1/HUWE1 control hemidesmosome maturation in C. elegans development. Current Biology 20 : 322–327.
56. CostanzoM, BaryshnikovaA, BellayJ, KimY, SpearED, et al. (2010) The genetic landscape of a cell. Science 327 : 425–431.
57. LiM, BrooksCL, KonN, GuW (2004) A dynamic role of HAUSP in the p53-Mdm2 pathway. Molecular cell 13 : 879–886.
58. ShengY, SaridakisV, SarkariF, DuanS, WuT, et al. (2006) Molecular recognition of p53 and MDM2 by USP7/HAUSP. Nature structural & molecular biology 13 : 285–291.
59. SongMS, SalmenaL, CarracedoA, EgiaA, Lo-CocoF, et al. (2008) The deubiquitinylation and localization of PTEN are regulated by a HAUSP-PML network. Nature 455 : 813–817.
60. BrajenovicM, JobertyG, KüsterB, BouwmeesterT, DrewesG (2004) Comprehensive proteomic analysis of human Par protein complexes reveals an interconnected protein network. The Journal of biological chemistry 279 : 12804–12811.
61. Al-HakimAK, ZagorskaA, ChapmanL, DeakM, PeggieM, et al. (2008) Control of AMPK-related kinases by USP9X and atypical Lys(29)/Lys(33)-linked polyubiquitin chains. The Biochemical journal 411 : 249–260.
62. CollandF, FormstecherE, JacqX, ReverdyC, PlanquetteC, et al. (2009) Small-molecule inhibitor of USP7/HAUSP ubiquitin protease stabilizes and activates p53 in cells. Molecular cancer therapeutics 8 : 2286–2295.
63. KowalskiJR, DahlbergCL, JuoP (2011) The deubiquitinating enzyme USP-46 negatively regulates the degradation of glutamate receptors to control their abundance in the ventral nerve cord of Caenorhabditis elegans. The Journal of Neuroscience 31 : 1341–1354.
64. BrennerS (1974) The Genetics of Caenorhabditis Elegans. Genetics 77 : 71–94.
65. GuoS, KemphuesKJ (1996) A non-muscle myosin required for embryonic polarity in Caenorhabditis elegans. Nature 382 : 455–458 Available:http://www.nature.com/nature/journal/v382/n6590/abs/382455a0.html.
66. KamathRS, FraserAG, DongY, PoulinG, DurbinR, et al. (2003) Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 421.
67. MaedaI, KoharaY, YamamotoM, SugimotoA (2001) Large-scale analysis of gene function in Caenorhabditis elegans by high-throughput RNAi. Current Biology 11 : 171–176.
68. RualJ-F, CeronJ, KorethJ, HaoT, NicotA-S, et al. (2004) Toward improving Caenorhabditis elegans phenome mapping with an ORFeome-based RNAi library. Genome research 14 : 2162–2168.
69. HurdDD, KemphuesKJ (2003) PAR-1 Is Required for Morphogenesis of the Caenorhabditis elegans Vulva. Developmental Biology 253 : 54–65.
70. TimmonsL, Firea (1998) Specific interference by ingested dsRNA. Nature 395 : 854.
71. CheesemanIM, NiessenS, AndersonS, HyndmanF, YatesJR, et al. (2004) A conserved protein network controls assembly of the outer kinetochore and its ability to sustain tension. Genes & development 18 : 2255–2268.
72. PraitisV, CaseyE, CollarD, AustinJ (2001) Creation of low-copy integrated transgenic lines in Caenorhabditis elegans. Genetics 157 : 1217–1226.
73. GattikerA, GasteigerE, BairochA (2002) ScanProsite: a reference implementation of a PROSITE scanning tool. Applied bioinformatics 1 : 107–108 Available:http://www.ncbi.nlm.nih.gov/pubmed/15130850.
74. FelsensteinJ (1989) PHYLIP – Phylogeny Inference Package (Version 3.2). Cladistics 5 : 163–166.
75. JohnsonM, ZaretskayaI, RaytselisY, MerezhukY, McGinnisS, et al. (2008) NCBI BLAST: a better web interface. Nucleic acids research 36: W5–9.
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