In macrophages, phagosome movement is microtubule-dependent. Microtubules are a prerequisite for phagosome maturation because they facilitate interactions between phagosomes and organelles of the endocytic pathway. We have established an in vitro assay that measures the binding of purified phagosomes to microtubules. This binding depends on the presence of membrane proteins, most likely integral to the surface of phagosomes, and on macrophage cytosol. The cytosolic binding factor can interact with microtubules prior to the addition of phagosomes to the assay, suggesting that it is a microtubule-associated protein (MAP). Consistent with this, depletion of MAPs from the cytosol by microtubule affinity removes all binding activity. Microtubule motor proteins show no binding activity, whereas a crude MAP preparation is sufficient to support binding and to restore full binding activity to MAP-depleted cytosol. We show that the activating MAP factor is a heat-sensitive protein(s) that migrates at around 150 kDa by gel filtration.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				M. Marsman, I. Jordens, C. Kuijl, L. Janssen, and J. Neefjes Dynein-mediated Vesicle Transport Controls Intracellular Salmonella Replication Mol. Biol. Cell, June 1, 2004; 15(6): 2954 - 2964. [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]

				C. Astarie-Dequeker, S. Carreno, C. Cougoule, and I. Maridonneau-Parini The protein tyrosine kinase Hck is located on lysosomal vesicles that are physically and functionally distinct from CD63-positive lysosomes in human macrophages J. Cell Sci., January 1, 2002; 115(1): 81 - 89. [Abstract] [Full Text] [PDF]

				A. Al-Haddad, M. A. Shonn, B. Redlich, A. Blocker, J. K. Burkhardt, H. Yu, J. A. Hammer III, D. G. Weiss, W. Steffen, G. Griffiths, et al. Myosin Va Bound to Phagosomes Binds to F-Actin and Delays Microtubule-dependent Motility Mol. Biol. Cell, September 1, 2001; 12(9): 2742 - 2755. [Abstract] [Full Text] [PDF]

				A. Naroeni, N. Jouy, S. Ouahrani-Bettache, J.-P. Liautard, and F. Porte Brucella suis-Impaired Specific Recognition of Phagosomes by Lysosomes due to Phagosomal Membrane Modifications Infect. Immun., January 1, 2001; 69(1): 486 - 493. [Abstract] [Full Text] [PDF]

				A Habermann, T. Schroer, G Griffiths, and J. Burkhardt Immunolocalization of cytoplasmic dynein and dynactin subunits in cultured macrophages: enrichment on early endocytic organelles J. Cell Sci., January 1, 2001; 114(1): 229 - 240. [Abstract] [PDF]

				A. Jahraus, M. Egeberg, B. Hinner, A. Habermann, E. Sackman, A. Pralle, H. Faulstich, V. Rybin, H. Defacque, and G. Griffiths ATP-dependent Membrane Assembly of F-Actin Facilitates Membrane Fusion Mol. Biol. Cell, January 1, 2001; 12(1): 155 - 170. [Abstract] [Full Text]

				I. Guerin and C. de Chastellier Pathogenic Mycobacteria Disrupt the Macrophage Actin Filament Network Infect. Immun., May 1, 2000; 68(5): 2655 - 2662. [Abstract] [Full Text] [PDF]

				J. Guttman, G. Kimel, and A. Vogl Dynein and plus-end microtubule-dependent motors are associated with specialized Sertoli cell junction plaques (ectoplasmic specializations) J. Cell Sci., January 6, 2000; 113(12): 2167 - 2176. [Abstract] [PDF]

				S. Timm, B. Titus, K. Bernd, and M. Barroso The EF-hand Ca2+-binding Protein p22 Associates with Microtubules in an N-Myristoylation-dependent Manner Mol. Biol. Cell, October 1, 1999; 10(10): 3473 - 3488. [Abstract] [Full Text]

				C. Infante, F. Ramos-Morales, C. Fedriani, M. Bornens, and R. M. Rios GMAP-210, A Cis-Golgi Network-associated Protein, Is a Minus End Microtubule-binding Protein J. Cell Biol., April 5, 1999; 145(1): 83 - 98. [Abstract] [Full Text] [PDF]

				A. Jahraus, T. E. Tjelle, T. Berg, A. Habermann, B. Storrie, O. Ullrich, and G. Griffiths In Vitro Fusion of Phagosomes with Different Endocytic Organelles from J774 Macrophages J. Biol. Chem., November 13, 1998; 273(46): 30379 - 30390. [Abstract] [Full Text] [PDF]

				E. J. PETTIT Cytosolic Free Calcium and the Cytoskeleton in the Control of Leukocyte Chemotaxis Physiol Rev, October 1, 1998; 78(4): 949 - 967. [Abstract] [Full Text] [PDF]

				S Searle, N. Bright, T. Roach, P. Atkinson, C. Barton, R. Meloen, and J. Blackwell Localisation of Nramp1 in macrophages: modulation with activation and infection J. Cell Sci., January 10, 1998; 111(19): 2855 - 2866. [Abstract] [PDF]

				A Blocker, G Griffiths, J. Olivo, A. Hyman, and F. Severin A role for microtubule dynamics in phagosome movement J. Cell Sci., January 2, 1998; 111(3): 303 - 312. [Abstract] [PDF]

				W. Steffen, S. Karki, K. T. Vaughan, R. B. Vallee, E. L.F. Holzbaur, D. G. Weiss, and S. A. Kuznetsov The Involvement of the Intermediate Chain of Cytoplasmic Dynein in Binding the Motor Complex to Membranous Organelles of Xenopus Oocytes Mol. Biol. Cell, October 1, 1997; 8(10): 2077 - 2088. [Abstract] [Full Text]

				K. Funato, W. Beron, C. Z. Yang, A. Mukhopadhyay, and P. D. Stahl Reconstitution of Phagosome-Lysosome Fusion in Streptolysin O-permeabilized Cells J. Biol. Chem., June 27, 1997; 272(26): 16147 - 16151. [Abstract] [Full Text] [PDF]

				A. Blocker, F. F. Severin, J. K. Burkhardt, J. B. Bingham, H. Yu, J.-C. Olivo, T. A. Schroer, A. A. Hyman, and G. Griffiths Molecular Requirements for Bi-directional Movement of Phagosomes Along Microtubules J. Cell Biol., April 7, 1997; 137(1): 113 - 129. [Abstract] [Full Text] [PDF]

				M Diakonova, V Gerke, J Ernst, J. Liautard, G van der Vusse, and G Griffiths Localization of five annexins in J774 macrophages and on isolated phagosomes J. Cell Sci., January 5, 1997; 110(10): 1199 - 1213. [Abstract] [PDF]

				C Alvarez-Dominguez, R Roberts, and P. Stahl Internalized Listeria monocytogenes modulates intracellular trafficking and delays maturation of the phagosome J. Cell Sci., January 3, 1997; 110(6): 731 - 743. [Abstract] [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]