Abstract
The position of lysosomes in the cytoplasm is important for their ability to fuse with the plasma membrane and release of proteases that are involved in tissue remodeling. Motor-directed bidirectional transport along microtubules is a critical process determining the distribution of lysosomes. How lysosomes are tethered to microtubules is incompletely understood, but a role for small GTPases of rab and arl families has been documented. We recently found that the rab5 and rab4 effector rabip4′ interacts with the adaptor complex AP-3 in a rab4-dependent manner on tubular endosomes. We here describe the assays that led to the identification of AP-3 as a rabip4′ partner and the role of the complex in regulating the spatial distribution of lysosomes.
An erratum to this chapter is available at http://dx.doi.org/10.1007/978-1-4939-2569-8_29
An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-4939-2569-8_29
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References
Luzio JP, Pryor PR, Bright NA (2007) Lysosomes: fusion and function. Nat Rev Mol Cell Biol 8:622–632
Settembre C, Fraldi A, Medina DL, Ballabio A (2013) Signals from the lysosome: a control centre for cellular clearance and energy metabolism. Nat Rev Mol Cell Biol 14:283–296
Marks MS, Heijnen HF, Raposo G (2013) Lysosome-related organelles: unusual compartments become mainstream. Curr Opin Cell Biol 25:495–505
Hendricks AG, Perlson E, Ross JL, Schroeder HW, Tokito M, Holzbauer ELF (2010) Motor coordination via a tug of war mechanism drives bidirectional vesicle transport. Curr Biol 20:697–702
Rosa-Ferreira C, Munro S (2011) Arl8 and SKIP act together to link lysosomes to kinesin-1. Dev Cell 21:1171–1178
Fouraux M, Deneka M, Ivan V, van der Heijden A, Raymackers J, van Suylekom D, van Venrooij WJ, van der Sluijs P, Pruijn GJM (2004) rabip4’ is an effector of rab5 and rab4 and regulates transport through early endosomes. Mol Biol Cell 15:611–624
Cormont M, Mari M, Galmiche A, Hofman P, Le Marchand-Brustel Y (2001) A FYVE finger-containing protein, rabip4, is a rab4 effector protein involved in early endosomal traffic. Proc Natl Acad Sci U S A 98:1637–1642
Theos AC, Tenza D, Martina JA, Hurbain I, Peden AA, Sviderskaya EV, Stewart A, Robinson MS, Bennett DC, Cutler DF et al (2005) Functions of adaptor protein AP-3 and AP-1 in tyrosinase sorting from endosomes to melanosomes. Mol Biol Cell 16:5356–5372
Peden AA, Oorschot V, Hesser BA, Austin CD, Scheller RH, Klumperman J (2004) Localization of the AP-3 adaptor complex defines a novel endosomal exit site for lysosomal membrane proteins. J Cell Biol 164:1065–1076
Dell’Angelica EC, Shotelersuk V, Aguilar RC, Gahl WA, Bonifacino JS (1999) Altered trafficking of lysosomal proteins in Hermansky-Pudlak syndrome due to mutations in the beta3A subunit of the AP-3 adaptor. Mol Cell 3:11–21
Clark RH, Stinchcombe JC, Day A, Blott E, Booth S, Bossi G, Hamblin T, Davies EG, Griffiths GM (2003) Adaptor protein 3-dependent microtubule mediated movement of lytic granules to the immunological synapse. Nat Immunol 4:1111–1120
Deneka M, Neeft M, Popa I, van Oort M, Sprong H, Oorschot V, Klumperman J, Schu P, van der Sluijs P (2003) rabaptin-5a/rabaptin-4 serves as a linker between rab4 and γ1-adaptin in membrane recycling from endosomes. EMBO J 22:2645–2657
Hoogenraad CC, Popa I, Futai K, Martinez-Sanchez E, Wulf PS, van Vlijmen T, Dortland B, Oorschot V, Govers R, Monti M et al (2010) Neuron specific rab4 effector GRASP-1 coordinates membrane specialization and maturation of recycling endosomes. PLoS Biol 8:e1000283
Yang J, Kim O, Qiu Y (2002) Interaction between tyrosine kinase Etk and a RUN domain and FYVE domain containing protein RUFY1. J Biol Chem 277:30219–30226
Yatsuda AP, Krijgsveld J, Cornelissen AW, Heck AJ, de Vries E (2003) Comprehensive analysis of the secreted proteins of the parasite Haemonchus contortus reveals extensive sequence variation and differential immune recognition. J Biol Chem 278:16941–16951
Ivan V, Martinez-Sanchez W, Sima LE, Oorschot V, Klumperman J, Petrescu SM, van der Sluijs P (2012) AP-3 and Rabip4' coordinately regulate spatial distribution of lysosomes. PLoS One 7:e48142
Acknowledgments
This research was supported by grant UEFISCDI PNII-RU 122/2010 (VI) and the Netherlands Organization for Scientific Research NWO-CW (PvdS).
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Ivan, V., van der Sluijs, P. (2015). Methods for Analysis of AP-3/Rabin4’ in Regulation of Lysosome Distribution. In: Li, G. (eds) Rab GTPases. Methods in Molecular Biology, vol 1298. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2569-8_21
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DOI: https://doi.org/10.1007/978-1-4939-2569-8_21
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