![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 12, Issue 1, 155-170, January 2001
*European Molecular Biology Laboratory, Heidelberg, Germany;
§Physik Department E22 Technische Universität
München, James Franck Strasse, 85747 Garching, Germany;
We recently established an in vitro assay that monitors the fusion
between latex-bead phagosomes and endocytic organelles in the presence
of J774 macrophage cytosol (Jahraus et al.,
1998). Here, we show that different reagents affecting the
actin cytoskeleton can either inhibit or stimulate this fusion process.
Because the membranes of purified phagosomes can assemble F-actin de
novo from pure actin with ATP (Defacque et al.,
2000a), we focused here on the ability of membranes to nucleate
actin in the presence of J774 cytosolic extracts. For this, we used
F-actin sedimentation, pyrene actin assays, and torsional rheometry, a
biophysical approach that could provide kinetic information on actin
polymerization and gel formation. We make two major conclusions. First,
under our standard in vitro conditions (4 mg/ml cytosol and 1 mM ATP), the presence of membranes actively catalyzed the assembly of cytosolic F-actin, which assembled into highly viscoelastic gels. A model is
discussed that links these results to how the actin may facilitate fusion. Second, cytosolic actin paradoxically polymerized more under
ATP depletion than under high-ATP conditions, even in the absence of
membranes; we discuss these data in the context of the well described,
large increases in F-actin seen in many cells during ischemia.
Max-Planck-Institut für Medizinische Forschung c/o
Max-Planck-Institut für Zell Biologie, Ladenburg, Germany
Present addresses: Laboratory
of Molecular Biology, Wageningen Agricultural University, Dreijenlaan
3, 6703 HA Wageningen, The Netherlands
¶
Present address: Observatorie Océanologique
de Banyuls, Laboratoire Arajo, 3P44, 66651 Sur Mer, France.
@
Corresponding author. European Molecular Biology
Laboratory, Meyerhoffstrasse 1, Postfach 102209, 69012 Heidelberg,
Germany. Fax: 0049 6221 387306. Tel: 0049 6221 387508. E-mail:
griffiths{at}embl-heidelberg.de.
These authors contributed equally to this study.
This article has been cited by other articles:
|
|
 |
|
  S. Isgandarova, L. Jones, D. Forsberg, A. Loncar, J. Dawson, K. Tedrick, and G. Eitzen Stimulation of Actin Polymerization by Vacuoles via Cdc42p-dependent Signaling J. Biol. Chem., October 19, 2007; 282(42): 30466 - 30475. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. St-Jean, T. Izard, and J. Sygusch A Hydrophobic Pocket in the Active Site of Glycolytic Aldolase Mediates Interactions with Wiskott-Aldrich Syndrome Protein J. Biol. Chem., May 11, 2007; 282(19): 14309 - 14315. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Trivedi, S. C. Zhang, A. B. Castoreno, W. Stockinger, E. C. Shieh, J. M. Vyas, E.-M. Frickel, and A. Nohturfft Immunoglobulin G signaling activates lysosome/phagosome docking PNAS, November 28, 2006; 103(48): 18226 - 18231. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. Kalamidas, M. P. Kuehnel, P. Peyron, V. Rybin, S. Rauch, O. B. Kotoulas, M. Houslay, B. A. Hemmings, M. G. Gutierrez, E. Anes, et al. cAMP synthesis and degradation by phagosomes regulate actin assembly and fusion events: consequences for mycobacteria. J. Cell Sci., September 1, 2006; 119(Pt 17): 3686 - 3694. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Burlak, A. R. Whitney, D. J. Mead, T. Hackstadt, and F. R. DeLeo Maturation of Human Neutrophil Phagosomes Includes Incorporation of Molecular Chaperones and Endoplasmic Reticulum Quality Control Machinery Mol. Cell. Proteomics, April 1, 2006; 5(4): 620 - 634. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Stockinger, S. C. Zhang, V. Trivedi, L. A. Jarzylo, E. C. Shieh, W. S. Lane, A. B. Castoreno, and A. Nohturfft Differential Requirements for Actin Polymerization, Calmodulin, and Ca2+ Define Distinct Stages of Lysosome/Phagosome Targeting Mol. Biol. Cell, April 1, 2006; 17(4): 1697 - 1710. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Tedrick, T. Trischuk, R. Lehner, and G. Eitzen Enhanced Membrane Fusion in Sterol-enriched Vacuoles Bypasses the Vrp1p Requirement Mol. Biol. Cell, October 1, 2004; 15(10): 4609 - 4621. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kanzaki, M. Furukawa, W. Raab, and J. E. Pessin Phosphatidylinositol 4,5-Bisphosphate Regulates Adipocyte Actin Dynamics and GLUT4 Vesicle Recycling J. Biol. Chem., July 16, 2004; 279(29): 30622 - 30633. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. A. Pfeiffer, M. Aga, U. Prabhu, J. J. Watters, D. J. Hall, and P. J. Bertics The nucleotide receptor P2X7 mediates actin reorganization and membrane blebbing in RAW 264.7 macrophages via p38 MAP kinase and Rho J. Leukoc. Biol., June 1, 2004; 75(6): 1173 - 1182. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. T. Watson, M. Kanzaki, and J. E. Pessin Regulated Membrane Trafficking of the Insulin-Responsive Glucose Transporter 4 in Adipocytes Endocr. Rev., April 1, 2004; 25(2): 177 - 204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. C. W. Richardson, S. C. Winistorfer, V. Poupon, J. P. Luzio, and R. C. Piper Mammalian Late Vacuole Protein Sorting Orthologues Participate in Early Endosomal Fusion and Interact with the Cytoskeleton Mol. Biol. Cell, March 1, 2004; 15(3): 1197 - 1210. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. J. Atkinson, M. A. Hosford, and B. A. Molitoris Mechanism of Actin Polymerization in Cellular ATP Depletion J. Biol. Chem., February 13, 2004; 279(7): 5194 - 5199. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Kjeken, M. Egeberg, A. Habermann, M. Kuehnel, P. Peyron, M. Floetenmeyer, P. Walther, A. Jahraus, H. Defacque, S. A. Kuznetsov, et al. Fusion between Phagosomes, Early and Late Endosomes: A Role for Actin in Fusion between Late, but Not Early Endocytic Organelles Mol. Biol. Cell, January 1, 2004; 15(1): 345 - 358. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. L. K. Hestvik, Z. Hmama, and Y. Av-Gay Kinome Analysis of Host Response to Mycobacterial Infection: a Novel Technique in Proteomics Infect. Immun., October 1, 2003; 71(10): 5514 - 5522. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Poupon, A. Stewart, S. R. Gray, R. C. Piper, and J. P. Luzio The Role of mVps18p in Clustering, Fusion, and Intracellular Localization of Late Endocytic Organelles Mol. Biol. Cell, October 1, 2003; 14(10): 4015 - 4027. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. S. Chew, X. Chen, J. A. Parente Jr, S. Tarrer, C. Okamoto, and H.-Y. Qin Lasp-1 binds to non-muscle F-actin in vitro and is localized within multiple sites of dynamic actin assembly in vivo J. Cell Sci., March 14, 2003; 115(24): 4787 - 4799. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Gotthardt, H. J. Warnatz, O. Henschel, F. Bruckert, M. Schleicher, and T. Soldati High-Resolution Dissection of Phagosome Maturation Reveals Distinct Membrane Trafficking Phases Mol. Biol. Cell, October 1, 2002; 13(10): 3508 - 3520. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Eitzen, L. Wang, N. Thorngren, and W. Wickner Remodeling of organelle-bound actin is required for yeast vacuole fusion J. Cell Biol., August 28, 2002; 158(4): 669 - 679. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Cougoule, P. Constant, G. Etienne, M. Daffe, and I. Maridonneau-Parini Lack of Fusion of Azurophil Granules with Phagosomes during Phagocytosis of Mycobacterium smegmatis by Human Neutrophils Is Not Actively Controlled by the Bacterium Infect. Immun., March 1, 2002; 70(3): 1591 - 1598. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Schratt, U. Philippar, J. Berger, H. Schwarz, O. Heidenreich, and A. Nordheim Serum response factor is crucial for actin cytoskeletal organization and focal adhesion assembly in embryonic stem cells J. Cell Biol., February 18, 2002; 156(4): 737 - 750. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Schratt, U. Philippar, J. Berger, H. Schwarz, O. Heidenreich, and A. Nordheim Serum response factor is crucial for actin cytoskeletal organization and focal adhesion assembly in embryonic stem cells J. Cell Biol., February 18, 2002; 156(4): 737 - 750. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Defacque, E. Bos, B. Garvalov, C. Barret, C. Roy, P. Mangeat, H.-W. Shin, V. Rybin, and G. Griffiths Phosphoinositides Regulate Membrane-dependent Actin Assembly by Latex Bead Phagosomes Mol. Biol. Cell, April 1, 2002; 13(4): 1190 - 1202. [Abstract] [Full Text] [PDF]
|
|
