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J Cell Biol
2002 Jan 21;1562:349-59. doi: 10.1083/jcb.200105051.
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Modulation of substrate adhesion dynamics via microtubule targeting requires kinesin-1.
Krylyshkina O
,
Kaverina I
,
Kranewitter W
,
Steffen W
,
Alonso MC
,
Cross RA
,
Small JV
.
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Recent studies have shown that the targeting of substrate adhesions by microtubules promotes adhesion site disassembly (Kaverina, I., O. Krylyshkina, and J.V. Small. 1999. J. Cell Biol. 146:1033-1043). It was accordingly suggested that microtubules serve to convey a signal to adhesion sites to modulate their turnover. Because microtubule motors would be the most likely candidates for effecting signal transmission, we have investigated the consequence of blocking microtubule motor activity on adhesion site dynamics. Using a function-blocking antibody as well as dynamitin overexpression, we found that a block in dynein-cargo interaction induced no change in adhesion site dynamics in Xenopus fibroblasts. In comparison, a block of kinesin-1 activity, either via microinjection of the SUK-4 antibody or of a kinesin-1 heavy chain construct mutated in the motor domain, induced a dramatic increase in the size and reduction in number of substrate adhesions, mimicking the effect observed after microtubule disruption by nocodazole. Blockage of kinesin activity had no influence on either the ability of microtubules to target substrate adhesions or on microtubule polymerisation dynamics. We conclude that conventional kinesin is not required for the guidance of microtubules into substrate adhesions, but is required for the focal delivery of a component(s) that retards their growth or promotes their disassembly.
Figure 1. Enlargement of focal adhesions following microtubule disassembly. Figure shows the same Xenopus fibroblast injected with TAMRA-vinculin, before (A) and after (B) treatment for 3 h with 2,5 μM nocodazole. (Video 8, available at http://www.jcb.org/cgi/content/full/jcb.200105051/DC1.)
Figure 2. Quantification of adhesion site size in control, nocodazole-treated and antibody-injected cells, as indicated. The data on adhesion site length was collected from image pairs of cells as in Fig. 1 (26 pairs for m74–2; 24 pairs for SUK-4 and 23 pairs for nocodazole), recorded immediately after injections (or beginning of treatment) and 3 h later. Note the closely similar increase in contact site size and decrease in number for nocodazole and kinesin inhibition with the SUK-4 antibody.
Figure 3. Inactivation of dynein by the m74–2 intermediate chain antibody, as monitored by the redistribution of lysosomes to the cell periphery. Lysosomes were loaded with rhodamine-dextran. (A and C) Paired phase contrast (A) and fluorescence (C) images of Xenopus fibroblast taken just after antibody injection. (B and D) The same cell 24 min later (Video 1, available at http://www.jcb.org/cgi/content/full/jcb.200105051/DC1).
Figure 4. Dynein inhibition has no effect on focal adhesions. Cells in A and C were injected with TAMRA vinculin to visualize adhesion sites. Cell in C was subsequently injected with the m74–2 anti-dynein antibody. After 3 h, the control (B) and antibody-injected cell (D) showed no enhancement of focal adhesions.
Figure 5. Control of block in kinesin motor activity by SUK-4 antibody. Figure shows living Xenopus fibroblasts in which mitochondria were marked with Rhodamine-123. The upper cell was injected with SUK-4 plus TAMRA dextran (diffuse background label in cytoplasm). Mitochondria are collapsed around the nucleus in the injected cell.
Figure 6. Induction and elongation of focal adhesions following inhibition of kinesin by the SUK-4 antibody. Two Xenopus fibroblasts are shown with adhesion sites marked with TAMRA vinculin (A and B) or GFP-zyxin (C and D). A and C show the cells just before injection of the SUK-4 antibody (0 min). B and D show the same cells respectively 3 h and 1 h later (Video 3, available at http://www.jcb.org/cgi/content/full/jcb.200105051/DC1).
Figure 7. Enlargement and elongation of focal adhesions following inhibition of kinesin by the rigor kinesin T93N. Two CAR fibroblasts are shown with adhesion sites marked with TAMRA vinculin (A and B) or GFP-zyxin (C and D). A and C show the cells just before injection of T93N kinesin (0 min). B and D show the same cells respectively 90 min and 1 h later (Video 4, available at http://www.jcb.org/cgi/content/full/jcb.200105051/DC1).
Figure 8. Microtubules distribution in Xenopus fibroblasts is unaffected by kinesin inhibition. Cells were preinjected with Cy-3–tubulin (control, middle) and additionally injected with 20 μM T93N (left) or SUK-4 antibody (right).
Figure 9. Microtubule targeting of substrate adhesions in Xenopus fibroblasts. Figure shows selected video frames from peripheral region of Xenopus fibroblasts coinjected with Cy-3–tubulin and TAMRA vinculin (Video 7, available at http://www.jcb.org/cgi/content/full/jcb.200105051/DC1). Arrows indicate targeting events. Time is given in minutes and seconds.
Figure 10. Kinesin inhibition does not block the targeting of substrate adhesions by microtubules. Figure shows video frames from a SUK-4 injected Xenopus fibroblast that was preinjected with Cy-3-tubulin and TAMRA-vinculin. Arrows indicate targeting events. Time is in minutes and seconds.
Akhmanova,
Clasps are CLIP-115 and -170 associating proteins involved in the regional regulation of microtubule dynamics in motile fibroblasts.
2001, Pubmed
Akhmanova,
Clasps are CLIP-115 and -170 associating proteins involved in the regional regulation of microtubule dynamics in motile fibroblasts.
2001,
Pubmed
Allan,
Catch and pull a microtubule: getting a grasp on the cortex.
2001,
Pubmed
Bershadsky,
Involvement of microtubules in the control of adhesion-dependent signal transduction.
1996,
Pubmed
Best,
The Ras-related GTPase Rac1 binds tubulin.
1996,
Pubmed
Bi,
Kinesin- and myosin-driven steps of vesicle recruitment for Ca2+-regulated exocytosis.
1997,
Pubmed
Brady,
A monoclonal antibody against kinesin inhibits both anterograde and retrograde fast axonal transport in squid axoplasm.
1990,
Pubmed
Browning,
Tea2p is a kinesin-like protein required to generate polarized growth in fission yeast.
2000,
Pubmed
Brunner,
CLIP170-like tip1p spatially organizes microtubular dynamics in fission yeast.
2000,
Pubmed
Burack,
Signal transduction: hanging on a scaffold.
2000,
Pubmed
Burkhardt,
Overexpression of the dynamitin (p50) subunit of the dynactin complex disrupts dynein-dependent maintenance of membrane organelle distribution.
1997,
Pubmed
Chrzanowska-Wodnicka,
Rho-stimulated contractility drives the formation of stress fibers and focal adhesions.
1996,
Pubmed
Danowski,
Microtubule dynamics in serum-starved and serum-stimulated Swiss 3T3 mouse fibroblasts: implications for the relationship between serum-induced contractility and microtubules.
1998,
Pubmed
Diamantopoulos,
Dynamic localization of CLIP-170 to microtubule plus ends is coupled to microtubule assembly.
1999,
Pubmed
Ebneth,
Overexpression of tau protein inhibits kinesin-dependent trafficking of vesicles, mitochondria, and endoplasmic reticulum: implications for Alzheimer's disease.
1998,
Pubmed
Enomoto,
Microtubule disruption induces the formation of actin stress fibers and focal adhesions in cultured cells: possible involvement of the rho signal cascade.
1996,
Pubmed
Garces,
Interaction of the p62 subunit of dynactin with Arp1 and the cortical actin cytoskeleton.
,
Pubmed
Gavin,
Microtubule-microfilament synergy in the cytoskeleton.
1997,
Pubmed
Glaven,
The Dbl-related protein, Lfc, localizes to microtubules and mediates the activation of Rac signaling pathways in cells.
1999,
Pubmed
Goode,
Functional cooperation between the microtubule and actin cytoskeletons.
2000,
Pubmed
Hirokawa,
Kinesin and dynein superfamily proteins and the mechanism of organelle transport.
1998,
Pubmed
Hollenbeck,
Radial extension of macrophage tubular lysosomes supported by kinesin.
1990,
Pubmed
Huang,
Direct interaction of microtubule- and actin-based transport motors.
1999,
Pubmed
Hunter,
How motor proteins influence microtubule polymerization dynamics.
2000,
Pubmed
Ingold,
Inhibition of kinesin-driven microtubule motility by monoclonal antibodies to kinesin heavy chains.
1988,
Pubmed
,
Xenbase
Kamal,
Connecting vesicle transport to the cytoskeleton.
2000,
Pubmed
Karki,
Cytoplasmic dynein and dynactin in cell division and intracellular transport.
1999,
Pubmed
Kaverina,
Enforced polarisation and locomotion of fibroblasts lacking microtubules.
2000,
Pubmed
Kaverina,
Kinesin-associated transport is involved in the regulation of cell adhesion.
1997,
Pubmed
Kaverina,
Targeting, capture, and stabilization of microtubules at early focal adhesions.
1998,
Pubmed
Kaverina,
Microtubule targeting of substrate contacts promotes their relaxation and dissociation.
1999,
Pubmed
Kawasaki,
Asef, a link between the tumor suppressor APC and G-protein signaling.
2000,
Pubmed
Kreitzer,
Kinesin and dynamin are required for post-Golgi transport of a plasma-membrane protein.
2000,
Pubmed
Lippincott-Schwartz,
Kinesin is the motor for microtubule-mediated Golgi-to-ER membrane traffic.
1995,
Pubmed
Lloyd,
Mechanisms of cellular adhesion. II. The interplay between adhesion, the cytoskeleton and morphology in substrate-attached cells.
1977,
Pubmed
Mata,
tea1 and the microtubular cytoskeleton are important for generating global spatial order within the fission yeast cell.
1997,
Pubmed
Mata,
Discovering the poles in yeast.
1998,
Pubmed
Mimori-Kiyosue,
Adenomatous polyposis coli (APC) protein moves along microtubules and concentrates at their growing ends in epithelial cells.
2000,
Pubmed
,
Xenbase
Mimori-Kiyosue,
The dynamic behavior of the APC-binding protein EB1 on the distal ends of microtubules.
2000,
Pubmed
,
Xenbase
Nagata,
The MAP kinase kinase kinase MLK2 co-localizes with activated JNK along microtubules and associates with kinesin superfamily motor KIF3.
1998,
Pubmed
Nakata,
Point mutation of adenosine triphosphate-binding motif generated rigor kinesin that selectively blocks anterograde lysosome membrane transport.
1995,
Pubmed
Näthke,
The adenomatous polyposis coli tumor suppressor protein localizes to plasma membrane sites involved in active cell migration.
1996,
Pubmed
Ren,
Cloning and characterization of GEF-H1, a microtubule-associated guanine nucleotide exchange factor for Rac and Rho GTPases.
1998,
Pubmed
Rinnerthaler,
Contact formation during fibroblast locomotion: involvement of membrane ruffles and microtubules.
1988,
Pubmed
Riveline,
Focal contacts as mechanosensors: externally applied local mechanical force induces growth of focal contacts by an mDia1-dependent and ROCK-independent mechanism.
2001,
Pubmed
Rodionov,
Microtubule-dependent control of cell shape and pseudopodial activity is inhibited by the antibody to kinesin motor domain.
1993,
Pubmed
Shelden,
Observation and quantification of individual microtubule behavior in vivo: microtubule dynamics are cell-type specific.
1993,
Pubmed
Smilenov,
Focal adhesion motility revealed in stationary fibroblasts.
1999,
Pubmed
Steffen,
Immunogold localisation of the intermediate chain within the protein complex of cytoplasmic dynein.
1996,
Pubmed
Steffen,
The involvement of the intermediate chain of cytoplasmic dynein in binding the motor complex to membranous organelles of Xenopus oocytes.
1997,
Pubmed
,
Xenbase
Terada,
Moving on to the cargo problem of microtubule-dependent motors in neurons.
2000,
Pubmed
Tuma,
Heterotrimeric kinesin II is the microtubule motor protein responsible for pigment dispersion in Xenopus melanophores.
1998,
Pubmed
,
Xenbase
Vaughan,
Colocalization of cytoplasmic dynein with dynactin and CLIP-170 at microtubule distal ends.
1999,
Pubmed
Verhey,
Cargo of kinesin identified as JIP scaffolding proteins and associated signaling molecules.
2001,
Pubmed
Wubbolts,
Opposing motor activities of dynein and kinesin determine retention and transport of MHC class II-containing compartments.
1999,
Pubmed
Zamir,
Dynamics and segregation of cell-matrix adhesions in cultured fibroblasts.
2000,
Pubmed
van Horck,
Characterization of p190RhoGEF, a RhoA-specific guanine nucleotide exchange factor that interacts with microtubules.
2001,
Pubmed
