QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Vol. 14, Issue 11, 4397-4413, November 2003

This Article Full Text Full Text (PDF) All Versions of this Article: E03-05-0315v1 14/11/4397    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by da Costa, S. R. Articles by Hamm-Alvarez, S. F. Search for Related Content PubMed PubMed Citation Articles by da Costa, S. R. Articles by Hamm-Alvarez, S. F.

Impairing Actin Filament or Syndapin Functions Promotes Accumulation of Clathrin-coated Vesicles at the Apical Plasma Membrane of Acinar Epithelial Cells

Silvia R. da Costa ^*, Eunbyul Sou ^*, Jiansong Xie , Francie A. Yarber ^*, Curtis T. Okamoto ^*, Michael Pidgeon , Michael M. Kessels ^||, Austin K. Mircheff  , Joel E. Schechter , Britta Qualmann ^||, and Sarah F. Hamm-Alvarez ^*   ¶

^* Department of Pharmaceutical Sciences, University of Southern California, Los Angeles, California 90033; Department of Physiology and Biophysics, University of Southern California, Los Angeles, California 90033; Department of Cell and Neurobiology, University of Southern California, Los Angeles, California 90033; Department of Ophthalmology, University of Southern California, Los Angeles, California 90033; and ^|| Leibniz Institute for Neurobiology, Magdeburg, Germany D-39118

Submitted May 19, 2003; Revised July 17, 2003; Accepted July 20, 2003
Monitoring Editor: Juan Bonifacino

In this article, we investigate the contributions of actin filaments and accessory proteins to apical clathrin-mediated endocytosis in primary rabbit lacrimal acini. Confocal fluorescence and electron microscopy revealed that cytochalasin D promoted apical accumulation of clathrin, -adaptin, dynamin, and F-actin and increased the amounts of coated pits and vesicles at the apical plasma membrane. Sorbitol density gradient analysis of membrane compartments showed that cytochalasin D increased [¹⁴C]dextran association with apical membranes from stimulated acini, consistent with functional inhibition of apical endocytosis. Recombinant syndapin SH3 domains interacted with lacrimal acinar dynamin, neuronal Wiskott-Aldrich Syndrome protein (N-WASP), and synaptojanin; their introduction by electroporation elicited remarkable accumulation of clathrin, accessory proteins, and coated pits at the apical plasma membrane. These SH3 domains also significantly (p 0.05) increased F-actin, with substantial colocalization of dynamin and N-WASP with the additional filaments. Coelectroporation with the VCA domain of N-WASP blocked the increase in F-actin and reversed the morphological changes indicative of impaired apical endocytosis. We suggest that transient modulation of actin polymerization by syndapins through activation of the Arp2/3 complex via N-WASP coordinates dynamin-mediated vesicle fission at the apical plasma membrane of acinar epithelia. Trapping of assembled F-actin intermediates during this process by cytochalasin D or syndapin SH3 domains impairs endocytosis.

^¶ Corresponding author. E-mail address: shalvar{at}usc.edu.

This article has been cited by other articles:

				R. R. Marchelletta, D. T. Jacobs, J. E. Schechter, R. E. Cheney, and S. F. Hamm-Alvarez The class V myosin motor, myosin 5c, localizes to mature secretory vesicles and facilitates exocytosis in lacrimal acini Am J Physiol Cell Physiol, July 1, 2008; 295(1): C13 - C28. [Abstract] [Full Text] [PDF]

				E. Evans, W. Zhang, G. Jerdeva, C.-Y. Chen, X. Chen, S. F. Hamm-Alvarez, and C. T. Okamoto Direct interaction between Rab3D and the polymeric immunoglobulin receptor and trafficking through regulated secretory vesicles in lacrimal gland acinar cells Am J Physiol Cell Physiol, March 1, 2008; 294(3): C662 - C674. [Abstract] [Full Text] [PDF]

				J. Xie, L. Chiang, J. Contreras, K. Wu, J. A. Garner, L. Medina-Kauwe, and S. F. Hamm-Alvarez Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in Lacrimal Acini J. Virol., December 1, 2006; 80(23): 11833 - 11851. [Abstract] [Full Text] [PDF]

				M. M. Kessels and B. Qualmann Syndapin Oligomers Interconnect the Machineries for Endocytic Vesicle Formation and Actin Polymerization J. Biol. Chem., May 12, 2006; 281(19): 13285 - 13299. [Abstract] [Full Text] [PDF]

				M. M. Kessels, J. Dong, W. Leibig, P. Westermann, and B. Qualmann Complexes of syndapin II with dynamin II promote vesicle formation at the trans-Golgi network J. Cell Sci., April 15, 2006; 119(8): 1504 - 1516. [Abstract] [Full Text] [PDF]

				G. V. Jerdeva, K. Wu, F. A. Yarber, C. J. Rhodes, D. Kalman, J. E. Schechter, and S. F. Hamm-Alvarez Actin and non-muscle myosin II facilitate apical exocytosis of tear proteins in rabbit lacrimal acinar epithelial cells J. Cell Sci., October 15, 2005; 118(20): 4797 - 4812. [Abstract] [Full Text] [PDF]

				G. V. Jerdeva, F. A. Yarber, M. D. Trousdale, C. J. Rhodes, C. T. Okamoto, D. A. Dartt, and S. F. Hamm-Alvarez Dominant-negative PKC-{epsilon} impairs apical actin remodeling in parallel with inhibition of carbachol-stimulated secretion in rabbit lacrimal acini Am J Physiol Cell Physiol, October 1, 2005; 289(4): C1052 - C1068. [Abstract] [Full Text] [PDF]

				S. Benesch, S. Polo, F. P. L. Lai, K. I. Anderson, T. E. B. Stradal, J. Wehland, and K. Rottner N-WASP deficiency impairs EGF internalization and actin assembly at clathrin-coated pits J. Cell Sci., July 15, 2005; 118(14): 3103 - 3115. [Abstract] [Full Text] [PDF]

				M. M. Kessels and B. Qualmann The syndapin protein family: linking membrane trafficking with the cytoskeleton J. Cell Sci., July 1, 2004; 117(15): 3077 - 3086. [Abstract] [Full Text] [PDF]

				A. M. Hommelgaard, M. Lerdrup, and B. van Deurs Association with Membrane Protrusions Makes ErbB2 an Internalization-resistant Receptor Mol. Biol. Cell, April 1, 2004; 15(4): 1557 - 1567. [Abstract] [Full Text] [PDF]