![]() |
|
|
Vol. 16, Issue 6, 2670-2680, June 2005
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
-Cells
* Henry Wellcome Laboratories for Integrated Cell Signalling and Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom;
Genomics Research Institute, Centre for Research in Biomedicine, University of the West of England, Bristol BS16 1QY, United Kingdom
Submitted November 16, 2004;
Revised February 7, 2005;
Accepted March 14, 2005
Monitoring Editor: Benjamin Glick
The role of unconventional myosins in neuroendocrine cells is not fully understood, with involvement suggested in the movement of both secretory vesicles and mitochondria. Here, we demonstrate colocalization of myosin Va (MyoVa) with insulin in pancreatic
-cells and show that MyoVa copurifies with insulin in density gradients and with the vesicle marker phogrin-enhanced green fluorescent protein upon fluorescence-activated sorting of vesicles. By contrast, MyoVa immunoreactivity was poorly colocalized with mitochondrial or other markers. Demonstrating an important role for MyoVa in the recruitment of secretory vesicles to the cell surface, a reduction of MyoVa protein levels achieved by RNA interference caused a significant decrease in glucose- or depolarization-stimulated insulin secretion. Similarly, expression of the dominant-negativeacting globular tail domain of MyoVa decreased by
50% the number of vesicles docked at the plasma membrane and by 87% the number of depolarization-stimulated exocytotic events detected by total internal reflection fluorescence microscopy. We conclude that MyoVa-driven movements of vesicles along the cortical actin network are essential for the terminal stages of regulated exocytosis in
-cells.
Abbreviations used: [Ca2+]i, intracellular free calcium ion concentration; ER, endoplasmic reticulum; EGFP, enhanced green fluorescent protein; FACS, fluorescence activated cell sorting; KRH, KrebsRinger-HEPES; LDCV, large dense core vesicle (secretory granule); MGTD, myosin Va globular tail domain; MT, microtubule; MyoVa, myosin Va; NPY, neuropeptide Y; siRNA, small interfering RNA; TIRF, total internal reflection fluorescence.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
Address correspondence to: Aniko Varadi (Aniko.Varadi{at}uwe.ac.uk) or Guy A. Rutter (g.a.rutter{at}bris.ac.uk).
This article has been cited by other articles:
![]() |
M. G. Pedersen and A. Sherman Newcomer insulin secretory granules as a highly calcium-sensitive pool PNAS, May 5, 2009; 106(18): 7432 - 7436. [Abstract] [Full Text] [PDF] |
||||
![]() |
S M Khoshnaw, A R Green, D G Powe, and I O Ellis MicroRNA involvement in the pathogenesis and management of breast cancer J. Clin. Pathol., May 1, 2009; 62(5): 422 - 428. [Abstract] [Full Text] [PDF] |
||||
![]() |
Z. Wang and D. C. Thurmond Mechanisms of biphasic insulin-granule exocytosis - roles of the cytoskeleton, small GTPases and SNARE proteins J. Cell Sci., April 1, 2009; 122(7): 893 - 903. [Abstract] [Full Text] [PDF] |
||||
![]() |
P. Bhat and P. Thorn Myosin 2 Maintains an Open Exocytic Fusion Pore in Secretory Epithelial Cells Mol. Biol. Cell, March 15, 2009; 20(6): 1795 - 1803. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. J. Park and Y. P. Loh Minireview: How Peptide Hormone Vesicles Are Transported to the Secretion Site for Exocytosis Mol. Endocrinol., December 1, 2008; 22(12): 2583 - 2595. [Abstract] [Full Text] [PDF] |
||||
![]() |
B. R. Gauthier and C. B. Wollheim Synaptotagmins bind calcium to release insulin Am J Physiol Endocrinol Metab, December 1, 2008; 295(6): E1279 - E1286. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. Trajkovski, H. Mziaut, S. Schubert, Y. Kalaidzidis, A. Altkruger, and M. Solimena Regulation of Insulin Granule Turnover in Pancreatic {beta}-Cells by Cleaved ICA512 J. Biol. Chem., November 28, 2008; 283(48): 33719 - 33729. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. H. Taft, F. K. Hartmann, A. Rump, H. Keller, I. Chizhov, D. J. Manstein, and G. Tsiavaliaris Dictyostelium Myosin-5b Is a Conditional Processive Motor J. Biol. Chem., October 3, 2008; 283(40): 26902 - 26910. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. L. Jewell, W. Luo, E. Oh, Z. Wang, and D. C. Thurmond Filamentous Actin Regulates Insulin Exocytosis through Direct Interaction with Syntaxin 4 J. Biol. Chem., April 18, 2008; 283(16): 10716 - 10726. [Abstract] [Full Text] [PDF] |
||||
![]() |
C. Desnos, S. Huet, I. Fanget, C. Chapuis, C. Bottiger, V. Racine, J.-B. Sibarita, J.-P. Henry, and F. Darchen Myosin Va Mediates Docking of Secretory Granules at the Plasma Membrane J. Neurosci., September 26, 2007; 27(39): 10636 - 10645. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Yoshizaki, T. Imamura, J. L. Babendure, J.-C. Lu, N. Sonoda, and J. M. Olefsky Myosin 5a Is an Insulin-Stimulated Akt2 (Protein Kinase B{beta}) Substrate Modulating GLUT4 Vesicle Translocation Mol. Cell. Biol., July 15, 2007; 27(14): 5172 - 5183. [Abstract] [Full Text] [PDF] |
||||
![]() |
S. Nofal, U. Becherer, D. Hof, U. Matti, and J. Rettig Primed Vesicles Can Be Distinguished from Docked Vesicles by Analyzing Their Mobility J. Neurosci., February 7, 2007; 27(6): 1386 - 1395. [Abstract] [Full Text] [PDF] |
||||
![]() |
A.-D. Lajoix, S. Badiou, S. Peraldi-Roux, T. Chardes, S. Dietz, C. Aknin, F. Tribillac, P. Petit, and R. Gross Protein Inhibitor of Neuronal Nitric Oxide Synthase (PIN) Is a New Regulator of Glucose-Induced Insulin Secretion Diabetes, December 1, 2006; 55(12): 3279 - 3288. [Abstract] [Full Text] [PDF] |
||||
![]() |
G. A. Rutter and E. V. Hill Insulin Vesicle Release: Walk, Kiss, Pause ... Then Run Physiology, June 1, 2006; 21(3): 189 - 196. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. W. Allersma, M. A. Bittner, D. Axelrod, and R. W. Holz Motion Matters: Secretory Granule Motion Adjacent to the Plasma Membrane and Exocytosis Mol. Biol. Cell, May 1, 2006; 17(5): 2424 - 2438. [Abstract] [Full Text] [PDF] |
||||
![]() |
P. E MacDonald, J. W Joseph, and P. Rorsman Glucose-sensing mechanisms in pancreatic {beta}-cells Phil Trans R Soc B, December 29, 2005; 360(1464): 2211 - 2225. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. Watanabe, K. Nomura, A. Ohyama, R. Ishikawa, Y. Komiya, K. Hosaka, E. Yamauchi, H. Taniguchi, N. Sasakawa, K. Kumakura, et al. Myosin-Va Regulates Exocytosis through the Submicromolar Ca2+-dependent Binding of Syntaxin-1A Mol. Biol. Cell, October 1, 2005; 16(10): 4519 - 4530. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. Krendel and M. S. Mooseker Myosins: Tails (and Heads) of Functional Diversity Physiology, August 1, 2005; 20(4): 239 - 251. [Abstract] [Full Text] [PDF] |
||||