Journal of Cell Science, Vol 107, Issue 2 683-692, Copyright © 1994 by Company of Biologists

JOURNAL ARTICLES

Tubulin and tektin in sea urchin embryonic cilia: pathways of protein incorporation during turnover and regeneration

RE Stephens
Marine Biological Laboratory, Woods Hole, MA 02543.

Axonemal precursor tubulin is the major protein component of the detergent-soluble membrane/matrix fraction of sea urchin embryonic cilia. Its unusual abundance may reflect the rapid turnover of these cilia, a process that is further documented here. However, whether during induced regeneration or normal turnover and growth, most other newly synthesized axonemal proteins are not detectable in the membrane/matrix fraction, raising the question of how non-tubulin precursors transit the growing cilium to the distal tip where assembly is generally thought to occur. Three potential explanations were considered: (1) the assembly of these components is proximal; (2) their relative concentration is too low to detect; or (3) tubulin alone is conveyed via a membrane/matrix pathway while most other axonemal proteins are transported in association with the axoneme. Light microscope autoradiography of axonemes pulse-chase labeled with [3H]leucine showed relatively uniform labeling, with no evidence for proximal incorporation. Fully grown cilia and cilia at early stages of regeneration were isolated from labeled embryos, fractionated into membrane/matrix, axonemal tubulin and architectural remnant components, and their labeled protein compositions were compared. Heavily labeled axonemal proteins, most notably the integral microtubule doublet component tektin-A, were not detected in the membrane/matrix fraction of emerging cilia, even though nearly half of the total ciliary tubulin appeared in that fraction, arguing against membrane-associated or soluble matrix transit for the architectural proteins at low concentrations. However, after thermal fractionation of axonemes from growing cilia, labeled proteins characteristic of the architectural remnant dominated the solubilized microtubule fraction, supporting axoneme-associated transport of the non-tubulin proteins during growth, in contrast to a membrane/matrix pathway for tubulin.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				A. I. Ivanov, I. C. McCall, C. A. Parkos, and A. Nusrat Role for Actin Filament Turnover and a Myosin II Motor in Cytoskeleton-driven Disassembly of the Epithelial Apical Junctional Complex Mol. Biol. Cell, June 1, 2004; 15(6): 2639 - 2651. [Abstract] [Full Text] [PDF]

				B. K. Straub, J. Boda, C. Kuhn, M. Schnoelzer, U. Korf, T. Kempf, H. Spring, M. Hatzfeld, and W. W. Franke A novel cell-cell junction system: the cortex adhaerens mosaic of lens fiber cells J. Cell Sci., December 15, 2003; 116(24): 4985 - 4995. [Abstract] [Full Text] [PDF]

				C. Casano, F. Gianguzza, M. C. Roccheri, R. Di Giorgi, L. Maenza, and M. A. Ragusa Hsp40 Is Involved in Cilia Regeneration in Sea Urchin Embryos J. Histochem. Cytochem., December 1, 2003; 51(12): 1581 - 1587. [Abstract] [Full Text] [PDF]

				N. Iguchi, H. Tanaka, Y. Nakamura, M. Nozaki, T. Fujiwara, and Y. Nishimune Cloning and characterization of the human tektin-t gene Mol. Hum. Reprod., June 1, 2002; 8(6): 525 - 530. [Abstract] [Full Text] [PDF]

				M. C. Chen, J. Goliger, N. Bunnett, and A. H. Soll Apical and basolateral EGF receptors regulate gastric mucosal paracellular permeability Am J Physiol Gastrointest Liver Physiol, February 1, 2001; 280(2): G264 - G272. [Abstract] [Full Text] [PDF]

				K Ersfeld and K Gull Targeting of cytoskeletal proteins to the flagellum of Trypanosoma brucei J. Cell Sci., January 1, 2001; 114(1): 141 - 148. [Abstract] [PDF]

				M. A. Jepson, H. B. Schlecht, and C. B. Collares-Buzato Localization of Dysfunctional Tight Junctions in Salmonella enterica Serovar Typhimurium-Infected Epithelial Layers Infect. Immun., December 1, 2000; 68(12): 7202 - 7208. [Abstract] [Full Text] [PDF]

				Y. Shi, M. Tabesh, and S. P. Sugrue Role of Cell Adhesion-Associated Protein, Pinin (DRS/memA), in Corneal Epithelial Migration Invest. Ophthalmol. Vis. Sci., May 1, 2000; 41(6): 1337 - 1345. [Abstract] [Full Text]

				M Rathman, P de Lanerolle, H Ohayon, P Gounon, and P Sansonetti Myosin light chain kinase plays an essential role in S. flexneri dissemination J. Cell Sci., January 10, 2000; 113(19): 3375 - 3386. [Abstract] [PDF]

				D. W. Owens, G. W. McLean, A. W. Wyke, C. Paraskeva, E. K. Parkinson, M. C. Frame, and V. G. Brunton The Catalytic Activity of the Src Family Kinases Is Required to Disrupt Cadherin-dependent Cell-Cell Contacts Mol. Biol. Cell, January 1, 2000; 11(1): 51 - 64. [Abstract] [Full Text]

				P. Bastin, T. H. MacRae, S. B. Francis, K. R. Matthews, and K. Gull Flagellar Morphogenesis: Protein Targeting and Assembly in the Paraflagellar Rod of Trypanosomes Mol. Cell. Biol., December 1, 1999; 19(12): 8191 - 8200. [Abstract] [Full Text] [PDF]

				R. Troyanovsky, J Klingelhofer, and S Troyanovsky Removal of calcium ions triggers a novel type of intercadherin interaction J. Cell Sci., January 12, 1999; 112(23): 4379 - 4387. [Abstract] [PDF]

				D. G. Cole, D. R. Diener, A. L. Himelblau, P. L. Beech, J. C. Fuster, and J. L. Rosenbaum Chlamydomonas Kinesin-II-dependent Intraflagellar Transport (IFT): IFT Particles Contain Proteins Required for Ciliary Assembly in Caenorhabditis elegans Sensory Neurons J. Cell Biol., May 18, 1998; 141(4): 993 - 1008. [Abstract] [Full Text] [PDF]

				M Perez-Moreno, A Avila, S Islas, S Sanchez, and L Gonzalez-Mariscal Vinculin but not alpha-actinin is a target of PKC phosphorylation during junctional assembly induced by calcium J. Cell Sci., January 12, 1998; 111(23): 3563 - 3571. [Abstract] [PDF]

				M. Balda and K Matter Tight junctions J. Cell Sci., January 3, 1998; 111(5): 541 - 547. [Abstract] [PDF]

				R. E. Stephens Synthesis and Turnover of Embryonic Sea Urchin Ciliary Proteins during Selective Inhibition of Tubulin Synthesis and Assembly Mol. Biol. Cell, November 1, 1997; 8(11): 2187 - 2198. [Abstract] [Full Text]

				C. Zhong, M. S. Kinch, and K. Burridge Rho-stimulated Contractility Contributes to the Fibroblastic Phenotype of Ras-transformed Epithelial Cells Mol. Biol. Cell, November 1, 1997; 8(11): 2329 - 2344. [Abstract] [Full Text]

				R. Gniadecki, B. Gajkowska, and M. Hansen 1,25-Dihydroxyvitamin D3 Stimulates the Assembly of Adherens Junctions in Keratinocytes: Involvement of Protein Kinase C Endocrinology, June 1, 1997; 138(6): 2241 - 2248. [Abstract] [Full Text] [PDF]

				F Monier-Gavelle and J. Duband Control of N-cadherin-mediated intercellular adhesion in migrating neural crest cells in vitro J. Cell Sci., January 12, 1995; 108(12): 3839 - 3853. [Abstract] [PDF]

				S Citi and N Denisenko Phosphorylation of the tight junction protein cingulin and the effects of protein kinase inhibitors and activators in MDCK epithelial cells J. Cell Sci., January 8, 1995; 108(8): 2917 - 2926. [Abstract] [PDF]

				J. Norrander, R. Linck, and R. Stephens Transcriptional control of tektin A mRNA correlates with cilia development and length determination during sea urchin embryogenesis Development, January 6, 1995; 121(6): 1615 - 1623. [Abstract] [PDF]

				J. Lewis, P. Jensen, K. Johnson, and M. Wheelock E-cadherin mediates adherens junction organization through protein kinase C J. Cell Sci., January 12, 1994; 107(12): 3615 - 3621. [Abstract] [PDF]

				N Denisenko, P Burighel, and S Citi Different effects of protein kinase inhibitors on the localization of junctional proteins at cell-cell contact sites J. Cell Sci., January 4, 1994; 107(4): 969 - 981. [Abstract] [PDF]