Export file:

Format

  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text

Content

  • Citation Only
  • Citation and Abstract

Non-centrosomal MTs play a crucial role in organization of MT array in interphase fibroblasts

1 Biology Department, M.V. Lomonosov Moscow State University, Moscow, Russia
2 A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Moscow, Russia
3 School of Science and Technology, and National Laboratory Astana, Nazarbayev University, Astana, Kazakhstan

Special Issues: Future of Biomedicine 2017

Microtubules in interphase fibroblast-like cells are thought to be organized in a radial array growing from a centrosome-based microtubule-organizing center (MTOC) to the cell edges. However, many morphogenetic processes require the asymmetry of the microtubules (MT) array. One of the possible mechanisms of this asymmetry could be the presence of non-centrosomal microtubules in different intracellular areas. To evaluate the role of centrosome-born and non-centrosomal microtubules in the organization of microtubule array in motile 3T3 fibroblasts, we have performed the high-throughput analysis of microtubule growth in different functional zones of the cell and distinguished three subpopulations of growing microtubules (centrosome-born, marginal and inner cytoplasmic).
Centrosome as an active microtubule-organizing center was absent in half of the cell population. However, these cells do not show any difference in microtubule growth pattern. In cells with active centrosome, it was constantly forming short (ephemeral) MTs, and ~15–20 MT per minute grow outwards for a distance >1 µm. Almost no persistent growth of microtubules was observed in these cells with the average growth length of 5–6 µm and duration of growth periods within 30 s.
However, the number of growing ends increased towards cell margin, especially towards the active edges. We found the peripheral cytoplasmic foci of microtubule growth there. During recovery from nocodazole treatment microtubules started to grow around the centrosome in a normal way and independently in all the cell areas. Within 5 minutes microtubules continued to grow mainly near the cell edge. Thus, our data confirm the negligible role of centrosome as MTOC in 3T3 fibroblasts and propose a model of non-centrosomal microtubules as major players that create the cell asymmetry in the cells with a mesenchymal type of motility. We suggest that increased density of dynamic microtubules near the active lamellum could be supported by microtubule-based microtubule nucleation.
  Figure/Table
  Article Metrics

Keywords cytoskeleton; centrosome; microtubule dynamics; non-centrosomal microtubules

Citation: Yekaterina Zvorykina, Anna Tvorogova, Aleena Gladkikh, Ivan Vorobjev. Non-centrosomal MTs play a crucial role in organization of MT array in interphase fibroblasts. AIMS Genetics, 2018, 5(2): 141-160. doi: 10.3934/genet.2018.2.141

References

  • 1. Rodionov V, Nadezhdina E, Borisy G (1999) Centrosomal control of microtubule dynamics. Proc Natl Acad Sci U.S.A 96: 115–120.    
  • 2. Nishita M, Satake T, Minami Y, et al. (2017) Regulatory mechanisms and cellular functions of non-centrosomal microtubules. J Biochem 162: 1–10.    
  • 3. Bartolini F, Gundersen GG (2006) Generation of noncentrosomal microtubule arrays. J Cell Sci 119: 4155–4163.    
  • 4. Irina BA, Tatyana B, Gary GB, et al. (2015) Centrosome nucleates numerous ephemeral microtubules and only few of them participate in the radial array. Cell Biol Int 39: 1203–1216.    
  • 5. Vorobjev IA, Chentsov YS (1983) The dynamics of reconstitution of microtubules around the cell center after cooling. Eur J Cell Biol 30: 149–153.
  • 6. Keating TJ, Peloquin JG, Rodionov VI, et al. (1997) Microtubule release from the centrosome. Proc Natl Acad Sci U.S.A 94: 5078–5083.    
  • 7. Waterman-Storer CM, Salmon ED (1997) Actomyosin-based retrograde flow of microtubules in the lamella of migrating epithelial cells influences microtubule dynamic instability and turnover and is associated with microtubule breakage and treadmilling. J Cell Sci 139: 417–434.    
  • 8. Vorobjev IA, Svitkina TM, Borisy GG (1997) Cytoplasmic assembly of microtubules in cultured cells. J Cell Sci 110: 2635–2645.
  • 9. Petry S, Vale RD (2015) Microtubule nucleation at the centrosome and beyond. Nat Cell Biol 17: 1089–1093.    
  • 10. Vorobjev IA, Alieva IB, Grigoriev IS, et al. (2003) Microtubule dynamics in living cells: Direct analysis in the internal cytoplasm. Cell Biol Int 27: 293–294.    
  • 11. Alieva IB, Borisy GG, Vorobjev IA (2008) Spatial organization of centrosome-attached and free microtubules in 3T3 fibroblasts. Cell Tissue Biol 50: 936–946.
  • 12. Komarova YA, Vorobjev IA, Borisy GG (2002) Life cycle of MTs: Persistent growth in the cell interior, asymmetric transition frequencies and effects of the cell boundary. J Cell Sci 115: 3527–3539.
  • 13. Vorobjev IA, Rodionov VI, Maly IV, et al. (1999) Contribution of plus and minus end pathways to microtubule turnover. J Cell Sci 112: 2277–2289.
  • 14. Komarova YA, Akhmanova AS, Kojima S, et al. (2002) Cytoplasmic linker proteins promote microtubule rescue in vivo. J Cell Biol 159: 589–599.    
  • 15. Vorob'Ev IA, Grigor'Ev IS, Borisy GG (2000) Microtubule dynamics in cultured cells. Ontogenez 31: 420–428.
  • 16. Matov A, Applegate K, Kumar P, et al. (2010) Analysis of microtubule dynamic instability using a plus-end growth marker. Nat Methods 7: 761–768.    
  • 17. Tanaka N, Meng W, Nagae S, et al. (2012) Nezha/CAMSAP3 and CAMSAP2 cooperate in epithelial-specific organization of noncentrosomal microtubules. Proc Natl Acad Sci U.S.A 109: 20029–20034.
  • 18. Efimov A, Kharitonov A, Efimova N, et al. (2007) Asymmetric CLASP-dependent nucleation of noncentrosomal microtubules at the trans-Golgi network. Dev Cell 12: 917–930.    
  • 19. Alieva IB, Zemskov EA, Kireev II, et al. (2010) Microtubules growth rate alteration in human endothelial cells. J Biomed Biotechnol 2010: 671536.
  • 20. Salaycik KJ, Fagerstrom CJ, Murthy K, et al. (2005) Quantification of microtubule nucleation, growth and dynamics in wound-edge cells. J Cell Sci 118: 4113–4122.    
  • 21. Gudima GO, Vorobjev IA, Chentsov YS (1988) Centriolar location during blood cell spreading and motion in vitro: An ultrastructural analysis. J Cell Sci 89: 225–241.
  • 22. Alieva IB, Vorobjev IA (1994) Centrosome behavior under the action of a mitochondrial uncoupler and the effect of disruption of cytoskeleton elements on the uncoupler-induced alterations. J Struct Biol 113: 217–224.    
  • 23. Komarova Y, De CG, Grigoriev I, et al. (2009) Mammalian end binding proteins control persistent microtubule growth. J Cell Biol 184: 691–706.    
  • 24. Li W, Moriwaki T, Tani T, et al. (2012) Reconstitution of dynamic microtubules with Drosophila XMAP215, EB1, and Sentin. J Cell Biol 199: 849–862.    
  • 25. Woodruff JB, Gomes BF, Widlund PO, et al. (2017) The Centrosome Is a Selective Condensate that Nucleates Microtubules by Concentrating Tubulin. Cell 169: 1066.    
  • 26. Wang L, Brown A (2002) Rapid movement of microtubules in axons. Curr Biol 12: 1496–1501.    
  • 27. Doxsey S, Mccollum D, Theurkauf W (2005) Centrosomes in cellular regulation. Annu Rev Cell Dev Biol 21: 411–434.    
  • 28. Rogers GC, Rusan NM, Peifer M, et al. (2008) A multicomponent assembly pathway contributes to the formation of acentrosomal microtubule arrays in interphase Drosophila cells. Mol Biol Cell 19: 3163–3178.    
  • 29. Luders J, Patel UK, Stearns T (2006) GCP-WD is a gamma-tubulin targeting factor required for centrosomal and chromatin-mediated microtubule nucleation. Nat Cell Biol 8: 137–147.    
  • 30. Cottam DM, Tucker JB, Rogers-Bald MM, et al. (2006) Non-centrosomal microtubule-organising centres in cold-treated cultured Drosophila cells. Cell Motil Cytoskeleton 63: 88–100.    
  • 31. Waterman-Storer CM, Worthylake RA, Liu BP, et al. (1999) Microtubule growth activates Rac1 to promote lamellipodial protrusion in fibroblasts. Nat Cell Biol 1: 45–50.    
  • 32. Rid R, Schiefermeier N, Grigoriev I, et al. (2005) The last but not the least: The origin and significance of trailing adhesions in fibroblastic cells. Cell Motil Cytoskeleton 61: 161–171.    
  • 33. Prigozhina NL, Waterman-Storer CM (2004) Protein kinase D-mediated anterograde membrane trafficking is required for fibroblast motility. Curr Biol 14: 88–98.    
  • 34. Shelden E, Wadsworth P (1993) Observation and quantification of individual microtubule behavior in vivo: Microtubule dynamics are cell-type specific. J Cell Biol 120: 935–945.    
  • 35. Chapin SJ, Bulinski JC (1991) Non-neuronal 210 × 103 Mr microtubule-associated protein (MAP4) contains a domain homologous to the microtubule-binding domains of neuronal MAP2 and tau. J Cell Sci 98: 27–36.
  • 36. Kinoshita K, Habermann B, Hyman AA (2002) XMAP215: A key component of the dynamic microtubule cytoskeleton. Trends Cell Biol 12: 267–273.    
  • 37. Toya M, Takeichi M (2016) Organization of Non-centrosomal Microtubules in Epithelial Cells. Cell Struct Funct 41: 127–135.    
  • 38. Guerin CM, Kramer SG (2009) Cytoskeletal remodeling during myotube assembly and guidance: Coordinating the actin and microtubule networks. Commun Integr Biol 2: 452–457.    
  • 39. Oddoux S, Zaal KJ, Tate V, et al. (2013) Microtubules that form the stationary lattice of muscle fibers are dynamic and nucleated at Golgi elements. J Cell Biol 203: 205–213.    
  • 40. Conde C, Caceres A (2009) Microtubule assembly, organization and dynamics in axons and dendrites. Nat Rev Neurosci 10: 319–332.    
  • 41. Akhmanova A, Steinmetz MO (2015) Control of microtubule organization and dynamics: Two ends in the limelight. Nat Rev Mol Cell Biol 16: 711–726.    
  • 42. Jiang K, Hua S, Mohan R, et al. (2014) Microtubule minus-end stabilization by polymerization-driven CAMSAP deposition. Dev Cell 28: 295–309.    
  • 43. Kamasaki T, O'Toole E, Kita S, et al. (2013) Augmin-dependent microtubule nucleation at microtubule walls in the spindle. J Cell Biol 202: 25–33.    
  • 44. Oakley BR, Paolillo V, Zheng Y (2015) Gamma-Tubulin complexes in microtubule nucleation and beyond. Mol Biol Cell 26: 2957–2962.    
  • 45. Uehara R, Nozawa RS, Tomioka A, et al. (2009) The augmin complex plays a critical role in spindle microtubule generation for mitotic progression and cytokinesis in human cells. Proc Natl Acad Sci U.S.A 106: 6998–7003.    
  • 46. Johmura Y, Soung NK, Park JE, et al. (2011) Regulation of microtubule-based microtubule nucleation by mammalian polo-like kinase 1. Proc Natl Acad Sci U.S.A 108: 11446–11451.    
  • 47. Hsia KC, Wilsonkubalek EM, Dottore A, et al. (2014) Reconstitution of the augmin complex provides insights into its architecture and function. Nat Cell Biol 16: 852–863.    

 

Copyright Info: © 2018, Ivan Vorobjev, licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

Download full text in PDF

Export Citation

Copyright © AIMS Press All Rights Reserved