ap-3 and rabip4’ coordinately regulate spatial distribution of lysosomesap-3和rabip4协调调节溶酶体的空间分布.pdfVIP

AP-3 and Rabip4’ Coordinately Regulate Spatial Distribution of Lysosomes 1,2 1 2 1 1 Viorica Ivan , Emma Martinez-Sanchez , Livia E. Sima , Viola Oorschot , Judith Klumperman , 2 1 Stefana M. Petrescu , Peter van der Sluijs * 1 Department of Cell Biology, University Medical Center Utrecht, Utrecht, The Netherlands, 2 Department of Molecular Cell Biology, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania Abstract The RUN and FYVE domain proteins rabip4 and rabip4’ are encoded by RUFY1 and differ in a 108 amino acid N-terminal extension in rabip4’. Their identical C terminus binds rab5 and rab4, but the function of rabip4s is incompletely understood. We here found that silencing RUFY1 gene products promoted outgrowth of plasma membrane protrusions, and polarized distribution and clustering of lysosomes at their tips. An interactor screen for proteins that function together with rabip4’ yielded the adaptor protein complex AP-3, of which the hinge region in the b3 subunit bound directly to the FYVE domain of rabip4’. Rabip4’ colocalized with AP-3 on a tubular subdomain of early endosomes and the extent of colocalization was increased by a dominant negative rab4 mutant. Knock-down of AP-3 had an ever more dramatic effect and caused accumulation of lysosomes in protrusions at the plasma membrane. The most peripheral lysosomes were localized beyond microtubules, within the cortical actin network. Our results uncover a novel function for AP-3 and rabip4’ in regulating lysosome positioning through an interorganellar pathway. Citation: Ivan V, Martinez-Sanchez E, Sima LE, Oorschot V, Klumperman J, et al.