Abstract
Polarized traffic in epithelial cells depends on well-organized pathways that direct secretory cargo to the apical or basolateral plasma membrane. In MDCK cells, apical trafficking can be further divided into a lipid raft-dependent and a raft-independent route, which separate biosynthetic cargo in a post-Golgi endosomal compartment. We have now identified KIF5C as a kinesin motor for apical trafficking of both raft-associated sucrase isomaltase and raft-independent neurotrophin receptor. KIF5C was identified by mass spectrometry in vesicle enriched fractions and on immunoisolated post-Golgi vesicles carrying apical cargo. The amount of vesicle-associated KIF5C was highest on material isolated directly after trans-Golgi network release and declined thereafter. Altogether, our data suggest that KIF5C is involved in the passage of apical cargo molecules to a post-Golgi endosomal compartment, where further segregation into distinct vesicle populations proceeds.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Rodriguez-Boulan E, Powell SK (1992) Polarity of epithelial and neuronal cells. Annu Rev Cell Biol 8:395–427
Danielsen EM, Hansen GH (2008) Lipid raft organization and function in the small intestinal brush border. J Physiol Biochem 64:377–382
Delacour D, Jacob R (2006) Apical protein transport. Cell Mol Life Sci 63:2491–2505
Griffiths G, Simons K (1986) The trans Golgi network: sorting at the exit site of the Golgi complex. Science 234:438–443
Jacob R, Naim HY (2001) Apical membrane proteins are transported in distinct vesicular carriers. Curr Biol 11:1444–1450
Jacob R, Heine M, Alfalah M, Naim HY (2003) Distinct cytoskeletal tracks direct individual vesicle populations to the apical membrane of epithelial cells. Curr Biol 13:607–612
Danielsen EM (1995) Involvement of detergent-insoluble complexes in the intracellular transport of intestinal brush border enzymes. Biochemistry 34:1596–1605
Fiedler K, Kobayashi T, Kurzchalia TV, Simons K (1993) Glycosphingolipid-enriched, detergent-insoluble complexes in protein sorting in epithelial cells. Biochemistry 32:6365–6373
Ojakian GK, Schwimmer R (1988) The polarized distribution of an apical cell surface glycoprotein is maintained by interactions with the cytoskeleton of Madin-Darby canine kidney cells. J Cell Biol 107:2377–2387
Zurzolo C, van’t Hof W, van Meer G, Rodriguez-Boulan E (1994) VIP21/caveolin, glycosphingolipid clusters and the sorting of glycosylphosphatidylinositol-anchored proteins in epithelial cells. EMBO J 13:42–53
Yeaman C, Le Gall AH, Baldwin AN, Monlauzeur L, Le Bivic A, Rodriguez-Boulan E (1997) The O-glycosylated stalk domain is required for apical sorting of neurotrophin receptors in polarized MDCK cells. J Cell Biol 139:929–940
Le Bivic A, Sambuy Y, Mostov K, Rodriguez-Boulan E (1990) Vectorial targeting of an endogenous apical membrane sialoglycoprotein and uvomorulin in MDCK cells. J Cell Biol 110:1533–1539
Verkade P, Harder T, Lafont F, Simons K (2000) Induction of caveolae in the apical plasma membrane of Madin-Darby canine kidney cells. J Cell Biol 148:727–739
Ihrke G, Bruns JR, Luzio JP, Weisz OA (2001) Competing sorting signals guide endolyn along a novel route to lysosomes in MDCK cells. EMBO J 20:6256–6264
Jacob R, Alfalah M, Grunberg J, Obendorf M, Naim HY (2000) Structural determinants required for apical sorting of an intestinal brush-border membrane protein. J Biol Chem 275:6566–6572
Musch A (2004) Microtubule organization and function in epithelial cells. Traffic 5:1–9
Bacallao R, Antony C, Dotti C, Karsenti E, Stelzer EH, Simons K (1989) The subcellular organization of Madin-Darby canine kidney cells during the formation of a polarized epithelium. J Cell Biol 109:2817–2832
Lafont F, Burkhardt JK, Simons K (1994) Involvement of microtubule motors in basolateral and apical transport in kidney cells. Nature 372:801–803
Noda Y, Okada Y, Saito N, Setou M, Xu Y, Zhang Z, Hirokawa N (2001) KIFC3, a microtubule minus end-directed motor for the apical transport of annexin XIIIb-associated Triton-insoluble membranes. J Cell Biol 155:77–88
Jaulin F, Xue X, Rodriguez-Boulan E, Kreitzer G (2007) Polarization-dependent selective transport to the apical membrane by KIF5B in MDCK cells. Dev Cell 13:511–522
Delacour D, Cramm-Behrens CI, Drobecq H, Le Bivic A, Naim HY, Jacob R (2006) Requirement for galectin-3 in apical protein sorting. Curr Biol 16:408–414
Cramm-Behrens CI, Dienst M, Jacob R (2008) Apical cargo traverses endosomal compartments on the passage to the cell surface. Traffic 9:2206–2220
Murray JW, Bananis E, Wolkoff AW (2000) Reconstitution of ATP-dependent movement of endocytic vesicles along microtubules in vitro: an oscillatory bidirectional process. Mol Biol Cell 11:419–433
Blocker A, Severin FF, Habermann A, Hyman AA, Griffiths G, Burkhardt JK (1996) Microtubule-associated protein-dependent binding of phagosomes to microtubules. J Biol Chem 271:3803–3811
Lu H, Ali MY, Bookwalter CS, Warshaw DM, Trybus KM (2009) Diffusive movement of processive kinesin-1 on microtubules. Traffic 10:1429–1438
Nangaku M, Sato-Yoshitake R, Okada Y, Noda Y, Takemura R, Yamazaki H, Hirokawa N (1994) KIF1B, a novel microtubule plus end-directed monomeric motor protein for transport of mitochondria. Cell 79:1209–1220
Wozniak MJ, Melzer M, Dorner C, Haring HU, Lammers R (2005) The novel protein KBP regulates mitochondria localization by interaction with a kinesin-like protein. BMC Cell Biol 6:35
Bohm KJ, Stracke R, Unger E (2000) Speeding up kinesin-driven microtubule gliding in vitro by variation of cofactor composition and physicochemical parameters. Cell Biol Int 24:335–341
Kanai Y, Okada Y, Tanaka Y, Harada A, Terada S, Hirokawa N (2000) KIF5C, a novel neuronal kinesin enriched in motor neurons. J Neurosci 20:6374–6384
Dathe V, Pröls F, Brand-Saberi B (2004) Expression of kinesin kif5c during chick development. Anat Embryol (Berl) 207:475–480
Smith MJ, Pozo K, Brickley K, Stephenson FA (2006) Mapping the GRIF-1 binding domain of the kinesin, KIF5C, substantiates a role for GRIF-1 as an adaptor protein in the anterograde trafficking of cargoes. J Biol Chem 281:27216–27228
Setou M, Seog DH, Tanaka Y, Kanai Y, Takei Y, Kawagishi M, Hirokawa N (2002) Glutamate-receptor-interacting protein GRIP1 directly steers kinesin to dendrites. Nature 417:83–87
Dunn S, Morrison EE, Liverpool TB, Molina-Paris C, Cross RA, Alonso MC, Peckham M (2008) Differential trafficking of Kif5c on tyrosinated and detyrosinated microtubules in live cells. J Cell Sci 121:1085–1095
Konishi Y, Setou M (2009) Tubulin tyrosination navigates the kinesin-1 motor domain to axons. Nat Neurosci 12:559–567
Cresawn KO, Potter BA, Oztan A, Guerriero CJ, Ihrke G, Goldenring JR, Apodaca G, Weisz OA (2007) Differential involvement of endocytic compartments in the biosynthetic traffic of apical proteins. EMBO J 26:3737–3748
Schmidt MR, Maritzen T, Kukhtina V, Higman VA, Doglio L, Barak NN, Strauss H, Oschkinat H, Dotti CG, Haucke V (2009) Regulation of endosomal membrane traffic by a Gadkin/AP-1/kinesin KIF5 complex. Proc Natl Acad Sci U S A 106:15344–15349
Cai Y, Singh BB, Aslanukov A, Zhao H, Ferreira PA (2001) The docking of kinesins, KIF5B and KIF5C, to Ran-binding protein 2 (RanBP2) is mediated via a novel RanBP2 domain. J Biol Chem 276:41594–41602
Acknowledgment
We are grateful to M. Dienst for technical assistance. Antibodies against SI and p75 were generous gifts from H.P. Hauri (Biocenter, University of Basel, Switzerland) and A. Le Bivic (Faculté des Sciences de Luminy, Marseille, France). This work was supported by the Deutsche Forschungsgemeinschaft (DFG), Bonn, Germany (grants JA 1033, Graduiertenkolleg 1216 and Sonderforschungsbereich 593).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Astanina, K., Jacob, R. KIF5C, a kinesin motor involved in apical trafficking of MDCK cells. Cell. Mol. Life Sci. 67, 1331–1342 (2010). https://doi.org/10.1007/s00018-009-0253-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00018-009-0253-6