TY - JOUR
T1 - Rapid whole cell imaging reveals a calcium-APPL1-dynein nexus that regulates cohort trafficking of stimulated EGF receptors
AU - York, H. M.
AU - Patil, A.
AU - Moorthi, U. K.
AU - Kaur, A.
AU - Bhowmik, A.
AU - Hyde, G. J.
AU - Gandhi, H.
AU - Fulcher, A.
AU - Gaus, K.
AU - Arumugam, S.
N1 - Funding Information:
This work was supported by the National Health and Medical Research Council of Australia (APP1182212) and ARC LIEF (LE150100163). H.M.Y. is supported by an Australian Government Research Training (RTP) Scholarship. A.P., U.K.M. are supported by Monash Biomedicine Discovery Institute Scholarships. The EMBL Australia Partnership Laboratory (EMBL Australia) is supported by the National Collaborative Research Infrastructure Strategy of the Australian Government. The authors would like to thank Profs. Joe Howard, Marino Zerial for engaging discussions and Drs. Vaishnavi Ananthanarayanan, Angika Basant, and Robert Weatheritt for valuable comments on the manuscript. The authors gratefully acknowledge the Imaging, FACS and Analysis Core and Cameron Nowell at Monash Institute of Pharmaceutical Science for their instrumentation and technical support. The authors acknowledge Monash Micro Imaging, Monash University, for the provision of instrumentation. The authors would like to thank the leadership at Faculty of Nursing, medicine and Health, Monash University, for COVID-safe access and facilitations to the microscope facilities made available to us through 2020 to be able to complete this manuscript.
Publisher Copyright:
© 2021, The Author(s).
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/2/17
Y1 - 2021/2/17
N2 - The endosomal system provides rich signal processing capabilities for responses elicited by growth factor receptors and their ligands. At the single cell level, endosomal trafficking becomes a critical component of signal processing, as exemplified by the epidermal growth factor (EGF) receptors. Activated EGFRs are trafficked to the phosphatase-enriched peri-nuclear region (PNR), where they are dephosphorylated and degraded. The details of the mechanisms that govern the movements of stimulated EGFRs towards the PNR, are not completely known. Here, exploiting the advantages of lattice light-sheet microscopy, we show that EGFR activation by EGF triggers a transient calcium increase causing a whole-cell level redistribution of Adaptor Protein, Phosphotyrosine Interacting with PH Domain And Leucine Zipper 1 (APPL1) from pre-existing endosomes within one minute, the rebinding of liberated APPL1 directly to EGFR, and the dynein-dependent translocation of APPL1-EGF-bearing endosomes to the PNR within ten minutes. The cell spanning, fast acting network that we reveal integrates a cascade of events dedicated to the cohort movement of activated EGF receptors. Our findings support the intriguing proposal that certain endosomal pathways have shed some of the stochastic strategies of traditional trafficking and have evolved processes that provide the temporal predictability that typify canonical signaling.
AB - The endosomal system provides rich signal processing capabilities for responses elicited by growth factor receptors and their ligands. At the single cell level, endosomal trafficking becomes a critical component of signal processing, as exemplified by the epidermal growth factor (EGF) receptors. Activated EGFRs are trafficked to the phosphatase-enriched peri-nuclear region (PNR), where they are dephosphorylated and degraded. The details of the mechanisms that govern the movements of stimulated EGFRs towards the PNR, are not completely known. Here, exploiting the advantages of lattice light-sheet microscopy, we show that EGFR activation by EGF triggers a transient calcium increase causing a whole-cell level redistribution of Adaptor Protein, Phosphotyrosine Interacting with PH Domain And Leucine Zipper 1 (APPL1) from pre-existing endosomes within one minute, the rebinding of liberated APPL1 directly to EGFR, and the dynein-dependent translocation of APPL1-EGF-bearing endosomes to the PNR within ten minutes. The cell spanning, fast acting network that we reveal integrates a cascade of events dedicated to the cohort movement of activated EGF receptors. Our findings support the intriguing proposal that certain endosomal pathways have shed some of the stochastic strategies of traditional trafficking and have evolved processes that provide the temporal predictability that typify canonical signaling.
UR - http://www.scopus.com/inward/record.url?scp=85100975482&partnerID=8YFLogxK
U2 - 10.1038/s42003-021-01740-y
DO - 10.1038/s42003-021-01740-y
M3 - Article
C2 - 33597720
AN - SCOPUS:85100975482
SN - 2399-3642
VL - 4
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 224
ER -