The polarity protein Dlg5 regulates collective cell migration during Drosophila oogenesis

Autoři: Jun Luo aff001;  Ping Zhou aff001;  Xuan Guo aff002;  Dou Wang aff003;  Jiong Chen aff002
Působiště autorů: College of Life Science, Shangrao Normal University, Shangrao, China aff001;  State Key Laboratory of Pharmaceutical Biotechnology and MOE Key Laboratory of Model Animals for Disease Study, Model Animal Research Center, Nanjing University, Nanjing, China aff002;  State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China aff003
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article


Collective migration plays critical roles in animal development, physiological events, and cancer metastasis. However, the molecular mechanisms of collective cell migration are not well understood. Drosophila border cells represent an excellent in vivo genetic model to study collective cell migration and identify novel regulatory genes for cell migration. Using the Mosaic Analysis with a Repressible Cell Marker (MARCM) system, we screened 240 P-element insertion lines to identify essential genes for border cell migration. Two genes were uncovered, including dlg5 (discs large 5) and CG31689. Further analysis showed that Dlg5 regulates the apical-basal polarity and cluster integrity in border cell clusters. Dlg5 is enriched in lateral surfaces between border cells and central polar cells but also shows punctate localization between border cells. We found that the distribution of Dlg5 in border cell clusters is regulated by Armadillo. Structure-function analysis revealed that the N-terminal Coiled-coil domain and the C-terminal PDZ3-PDZ4-SH3-GUK domains but not the PDZ1-PDZ2 domains of Dlg5 are required for BC migration. The Coiled-coil domain and the PDZ4-SH3-GUK domains are critical for Dlg5’s cell surface localization in border cell clusters.

Klíčová slova:

Cell migration – Cell polarity – Drosophila melanogaster – Hyperexpression techniques – RNA interference – Subcellular localization – Parietal cells


1. Friedl P, Gilmour D. Collective cell migration in morphogenesis, regeneration and cancer. Nature reviews Molecular cell biology. 2009;10(7):445–57. doi: 10.1038/nrm2720 19546857.

2. Yilmaz M, Christofori G. Mechanisms of motility in metastasizing cells. Molecular cancer research: MCR. 2010;8(5):629–42. doi: 10.1158/1541-7786.MCR-10-0139 20460404.

3. Friedl P, Locker J, Sahai E, Segall JE. Classifying collective cancer cell invasion. Nature cell biology. 2012;14(8):777–83. doi: 10.1038/ncb2548 22854810.

4. He L, Wang X, Montell DJ. Shining light on Drosophila oogenesis: live imaging of egg development. Current opinion in genetics & development. 2011;21(5):612–9. doi: 10.1016/j.gde.2011.08.011 21930372.

5. Montell DJ. Border-cell migration: the race is on. Nature reviews Molecular cell biology. 2003;4(1):13–24. Epub 2003/01/04. doi: 10.1038/nrm1006 12511865.

6. Montell DJ, Rorth P, Spradling AC. slow border cells, a locus required for a developmentally regulated cell migration during oogenesis, encodes Drosophila C/EBP. Cell. 1992;71(1):51–62. Epub 1992/10/02. doi: 10.1016/0092-8674(92)90265-e 1394432.

7. McDonald JA, Pinheiro EM, Montell DJ. PVF1, a PDGF/VEGF homolog, is sufficient to guide border cells and interacts genetically with Taiman. Development. 2003;130(15):3469–78. doi: 10.1242/dev.00574 12810594.

8. Starz-Gaiano M, Melani M, Wang X, Meinhardt H, Montell DJ. Feedback inhibition of Jak/STAT signaling by apontic is required to limit an invasive cell population. Dev Cell. 2008;14(5):726–38. doi: 10.1016/j.devcel.2008.03.005 18477455.

9. McDonald JA, Khodyakova A, Aranjuez G, Dudley C, Montell DJ. PAR-1 kinase regulates epithelial detachment and directional protrusion of migrating border cells. Current biology: CB. 2008;18(21):1659–67. doi: 10.1016/j.cub.2008.09.041 18976916; PubMed Central PMCID: PMC2593744.

10. Bai J, Uehara Y, Montell DJ. Regulation of invasive cell behavior by taiman, a Drosophila protein related to AIB1, a steroid receptor coactivator amplified in breast cancer. Cell. 2000;103(7):1047–58. Epub 2001/02/13. doi: 10.1016/s0092-8674(00)00208-7 11163181.

11. Liu Y, Montell DJ. Jing: a downstream target of slbo required for developmental control of border cell migration. Development. 2001;128(3):321–30. Epub 2001/01/12. 11152631.

12. Kim JH, Cho A, Yin H, Schafer DA, Mouneimne G, Simpson KJ, et al. Psidin, a conserved protein that regulates protrusion dynamics and cell migration. Genes & development. 2011;25(7):730–41. Epub 2011/03/17. doi: 10.1101/gad.2028611 21406550; PubMed Central PMCID: PMC3070935.

13. Jang AC, Chang YC, Bai J, Montell D. Border-cell migration requires integration of spatial and temporal signals by the BTB protein Abrupt. Nature cell biology. 2009;11(5):569–79. Epub 2009/04/08. doi: 10.1038/ncb1863 19350016; PubMed Central PMCID: PMC2675665.

14. Llense F, Martin-Blanco E. JNK signaling controls border cell cluster integrity and collective cell migration. Current biology: CB. 2008;18(7):538–44. Epub 2008/04/09. doi: 10.1016/j.cub.2008.03.029 18394890.

15. Wan P, Wang D, Luo J, Chu D, Wang H, Zhang L, et al. Guidance receptor promotes the asymmetric distribution of exocyst and recycling endosome during collective cell migration. Development. 2013;140(23):4797–806. Epub 2013/11/08. doi: 10.1242/dev.094979 24198275.

16. Pinheiro EM, Montell DJ. Requirement for Par-6 and Bazooka in Drosophila border cell migration. Development. 2004;131(21):5243–51. Epub 2004/10/01. doi: 10.1242/dev.01412 15456726.

17. Niewiadomska P, Godt D, Tepass U. DE-Cadherin is required for intercellular motility during Drosophila oogenesis. The Journal of cell biology. 1999;144(3):533–47. Epub 1999/02/11. doi: 10.1083/jcb.144.3.533 9971747; PubMed Central PMCID: PMC2132905.

18. Goode S, Perrimon N. Inhibition of patterned cell shape change and cell invasion by Discs large during Drosophila oogenesis. Genes & development. 1997;11(19):2532–44. Epub 1997/10/23. doi: 10.1101/gad.11.19.2532 9334318; PubMed Central PMCID: PMC316565.

19. Goode S, Wei J, Kishore S. Novel spatiotemporal patterns of epithelial tumor invasion in Drosophila discs large egg chambers. Developmental dynamics: an official publication of the American Association of Anatomists. 2005;232(3):855–64. Epub 2005/02/16. doi: 10.1002/dvdy.20336 15712204.

20. Wang H, Qiu Z, Xu Z, Chen SJ, Luo J, Wang X, et al. aPKC is a key polarity determinant in coordinating the function of three distinct cell polarities during collective migration. Development. 2018;145(9). doi: 10.1242/dev.158444 29636381.

21. Bellen HJ, Levis RW, Liao G, He Y, Carlson JW, Tsang G, et al. The BDGP gene disruption project: single transposon insertions associated with 40% of Drosophila genes. Genetics. 2004;167(2):761–81. doi: 10.1534/genetics.104.026427 15238527; PubMed Central PMCID: PMC1470905.

22. Theodosiou NA, Xu T. Use of FLP/FRT system to study Drosophila development. Methods. 1998;14(4):355–65. Epub 1998/06/03. doi: 10.1006/meth.1998.0591 9608507.

23. Perrimon N. Creating mosaics in Drosophila. The International journal of developmental biology. 1998;42(3):243–7. Epub 1998/07/08. 9654004.

24. Lee T, Luo L. Mosaic analysis with a repressible cell marker (MARCM) for Drosophila neural development. Trends in neurosciences. 2001;24(5):251–4. doi: 10.1016/s0166-2236(00)01791-4 11311363.

25. Chen J, Call GB, Beyer E, Bui C, Cespedes A, Chan A, et al. Discovery-based science education: functional genomic dissection in Drosophila by undergraduate researchers. PLoS biology. 2005;3(2):e59. Epub 2005/02/19. doi: 10.1371/journal.pbio.0030059 15719063; PubMed Central PMCID: PMC548953.

26. te Velthuis AJ, Admiraal JF, Bagowski CP. Molecular evolution of the MAGUK family in metazoan genomes. BMC evolutionary biology. 2007;7:129. Epub 2007/08/07. doi: 10.1186/1471-2148-7-129 17678554; PubMed Central PMCID: PMC1978500.

27. Nechiporuk T, Fernandez TE, Vasioukhin V. Failure of epithelial tube maintenance causes hydrocephalus and renal cysts in Dlg5-/- mice. Dev Cell. 2007;13(3):338–50. Epub 2007/09/04. doi: 10.1016/j.devcel.2007.07.017 17765678.

28. Nakamura H, Sudo T, Tsuiki H, Miyake H, Morisaki T, Sasaki J, et al. Identification of a novel human homolog of the Drosophila dlg, P-dlg, specifically expressed in the gland tissues and interacting with p55. FEBS letters. 1998;433(1–2):63–7. Epub 1998/09/17. doi: 10.1016/s0014-5793(98)00882-5 9738934.

29. Wakabayashi M, Ito T, Mitsushima M, Aizawa S, Ueda K, Amachi T, et al. Interaction of lp-dlg/KIAA0583, a membrane-associated guanylate kinase family protein, with vinexin and beta-catenin at sites of cell-cell contact. The Journal of biological chemistry. 2003;278(24):21709–14. Epub 2003/03/27. doi: 10.1074/jbc.M211004200 12657639.

30. Chang Y, Klezovitch O, Walikonis RS, Vasioukhin V, LoTurco JJ. Discs large 5 is required for polarization of citron kinase in mitotic neural precursors. Cell Cycle. 2010;9(10):1990–7. Epub 2010/05/04. doi: 10.4161/cc.9.10.11730 20436275.

31. Sezaki T, Tomiyama L, Kimura Y, Ueda K, Kioka N. Dlg5 interacts with the TGF-beta receptor and promotes its degradation. FEBS letters. 2013;587(11):1624–9. Epub 2013/04/30. doi: 10.1016/j.febslet.2013.04.015 23624079.

32. Tomiyama L, Sezaki T, Matsuo M, Ueda K, Kioka N. Loss of Dlg5 expression promotes the migration and invasion of prostate cancer cells via Girdin phosphorylation. Oncogene. 2015;34(9):1141–9. doi: 10.1038/onc.2014.31 24662825.

33. Chong YC, Mann RK, Zhao C, Kato M, Beachy PA. Bifurcating action of Smoothened in Hedgehog signaling is mediated by Dlg5. Genes & development. 2015;29(3):262–76. doi: 10.1101/gad.252676.114 25644602; PubMed Central PMCID: PMC4318143.

34. Nechiporuk T, Klezovitch O, Nguyen L, Vasioukhin V. Dlg5 maintains apical aPKC and regulates progenitor differentiation during lung morphogenesis. Developmental biology. 2013;377(2):375–84. Epub 2013/03/08. doi: 10.1016/j.ydbio.2013.02.019 23466739; PubMed Central PMCID: PMC3652546.

35. Aranjuez G, Kudlaty E, Longworth MS, McDonald JA. On the role of PDZ domain-encoding genes in Drosophila border cell migration. G3 (Bethesda). 2012;2(11):1379–91. Epub 2012/11/23. doi: 10.1534/g3.112.004093 23173089.

36. Reilly E, Changela N, Naryshkina T, Deshpande G, Steward R. Discs large 5, an Essential Gene in Drosophila, Regulates Egg Chamber Organization. G3 (Bethesda). 2015. doi: 10.1534/g3.115.017558 25795662.

37. Luo J, Wang H, Kang D, Guo X, Wan P, Wang D, et al. Dlg5 maintains apical polarity by promoting membrane localization of Crumbs during Drosophila oogenesis. Scientific reports. 2016;6:26553. doi: 10.1038/srep26553 27211898; PubMed Central PMCID: PMC4876392.

38. Huang J, Zhou W, Dong W, Watson AM, Hong Y. From the Cover: Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering. Proceedings of the National Academy of Sciences of the United States of America. 2009;106(20):8284–9. Epub 2009/05/12. doi: 10.1073/pnas.0900641106 19429710; PubMed Central PMCID: PMC2688891.

39. Doerflinger H, Vogt N, Torres IL, Mirouse V, Koch I, Nusslein-Volhard C, et al. Bazooka is required for polarisation of the Drosophila anterior-posterior axis. Development. 2010;137(10):1765–73. Epub 2010/05/01. doi: 10.1242/dev.045807 20430751; PubMed Central PMCID: PMC2860254.

40. Lee CY, Robinson KJ, Doe CQ. Lgl, Pins and aPKC regulate neuroblast self-renewal versus differentiation. Nature. 2006;439(7076):594–8. Epub 2005/12/17. doi: 10.1038/nature04299 16357871.

41. Ghiglione C, Devergne O, Cerezo D, Noselli S. Drosophila RalA is essential for the maintenance of Jak/Stat signalling in ovarian follicles. EMBO reports. 2008;9(7):676–82. doi: 10.1038/embor.2008.79 18552769; PubMed Central PMCID: PMC2475328.

42. Luo J, Shen P, Chen J. A modular toolset of phiC31-based fluorescent protein tagging vectors for Drosophila. Fly. 2019:1–13. doi: 10.1080/19336934.2019.1664363 30885036.

43. Call GB, Olson JM, Chen J, Villarasa N, Ngo KT, Yabroff AM, et al. Genomewide clonal analysis of lethal mutations in the Drosophila melanogaster eye: comparison of the X chromosome and autosomes. Genetics. 2007;177(2):689–97. doi: 10.1534/genetics.107.077735 17720911; PubMed Central PMCID: PMC2034635.

44. Kadrmas JL, Smith MA, Pronovost SM, Beckerle MC. Characterization of RACK1 function in Drosophila development. Developmental dynamics: an official publication of the American Association of Anatomists. 2007;236(8):2207–15. doi: 10.1002/dvdy.21217 17584887.

45. Luo J, Zuo J, Wu J, Wan P, Kang D, Xiang C, et al. In vivo RNAi screen identifies candidate signaling genes required for collective cell migration in Drosophila ovary. Science China Life sciences. 2015;58(4):379–89. doi: 10.1007/s11427-014-4786-z 25528253.

46. Ando R, Mizuno H, Miyawaki A. Regulated fast nucleocytoplasmic shuttling observed by reversible protein highlighting. Science. 2004;306(5700):1370–3. doi: 10.1126/science.1102506 15550670.

47. Anderson JM. Cell signalling: MAGUK magic. Current biology: CB. 1996;6(4):382–4. Epub 1996/04/01. doi: 10.1016/s0960-9822(02)00501-8 8723338.

48. Gonzalez-Mariscal L, Betanzos A, Avila-Flores A. MAGUK proteins: structure and role in the tight junction. Seminars in cell & developmental biology. 2000;11(4):315–24. Epub 2000/09/01. doi: 10.1006/scdb.2000.0178 10966866.

49. Caruana G. Genetic studies define MAGUK proteins as regulators of epithelial cell polarity. The International journal of developmental biology. 2002;46(4):511–8. Epub 2002/07/27. 12141438.

50. Funke L, Dakoji S, Bredt DS. Membrane-associated guanylate kinases regulate adhesion and plasticity at cell junctions. Annual review of biochemistry. 2005;74:219–45. Epub 2005/06/15. doi: 10.1146/annurev.biochem.74.082803.133339 15952887.

51. Dietzl G, Chen D, Schnorrer F, Su KC, Barinova Y, Fellner M, et al. A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature. 2007;448(7150):151–6. doi: 10.1038/nature05954 17625558.

52. Manseau L, Baradaran A, Brower D, Budhu A, Elefant F, Phan H, et al. GAL4 enhancer traps expressed in the embryo, larval brain, imaginal discs, and ovary of Drosophila. Developmental dynamics: an official publication of the American Association of Anatomists. 1997;209(3):310–22. Epub 1997/07/01. doi: 10.1002/(SICI)1097-0177(199707)209:3<310::AID-AJA6>3.0.CO;2-L 9215645.

53. Matthews KA, Miller DF, Kaufman TC. Developmental distribution of RNA and protein products of the Drosophila alpha-tubulin gene family. Developmental biology. 1989;132(1):45–61. doi: 10.1016/0012-1606(89)90203-0 2492961.

54. Jia D, Xu Q, Xie Q, Mio W, Deng WM. Automatic stage identification of Drosophila egg chamber based on DAPI images. Scientific reports. 2016;6:18850. doi: 10.1038/srep18850 26732176; PubMed Central PMCID: PMC4702167.

55. Tanentzapf G, Smith C, McGlade J, Tepass U. Apical, lateral, and basal polarization cues contribute to the development of the follicular epithelium during Drosophila oogenesis. The Journal of cell biology. 2000;151(4):891–904. Epub 2000/11/15. doi: 10.1083/jcb.151.4.891 11076972; PubMed Central PMCID: PMC2169434.

56. Taniuchi K, Nakagawa H, Nakamura T, Eguchi H, Ohigashi H, Ishikawa O, et al. Down-regulation of RAB6KIFL/KIF20A, a kinesin involved with membrane trafficking of discs large homologue 5, can attenuate growth of pancreatic cancer cell. Cancer research. 2005;65(1):105–12. Epub 2005/01/25. 15665285.

57. Smolen GA, Zhang J, Zubrowski MJ, Edelman EJ, Luo B, Yu M, et al. A genome-wide RNAi screen identifies multiple RSK-dependent regulators of cell migration. Genes & development. 2010;24(23):2654–65. Epub 2010/11/11. doi: 10.1101/gad.1989110 21062900; PubMed Central PMCID: PMC2994039.

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