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Publication details

Document type
Journal articles

Document subtype
Full paper

Title
A novel microscopy-based assay identifies extended synaptotagmin-1 (ESYT1) as a positive regulator of anoctamin 1 traffic

Participants in the publication
Joana R. Lérias (Author)
BioISI - Biosystems & Integrative Sciences Institute
Madalena C. Pinto (Author)
BIOISI - Instituto de Biossistemas e Ciências Integrativas
Hugo M. Botelho (Author)
Nikhil T. Awatade (Author)
Margarida C. Quaresma (Author)
Iris A.L. Silva (Author)
BIOISI - Instituto de Biossistemas e Ciências Integrativas
Podchanart Wanitchakool (Author)
Rainer Schreiber (Author)
Rainer Pepperkok (Author)
Karl Kunzelmann (Author)
Margarida D. Amaral (Author)
Dep. Química e Bioquímica
BIOISI - Instituto de Biossistemas e Ciências Integrativas

Scope
International

Refereeing
Yes

Summary
An attractive possibility to treat Cystic Fibrosis (CF), a severe condition caused by dysfunctional CFTR, an epithelial anion channel, is through the activation of alternative (non-CFTR) anion channels. Anoctamin 1 (ANO1) was demonstrated to be a Ca2+-activated chloride channel (CaCC) and thus of high potential to replace CFTR. Despite that ANO1 is expressed in human lung CF tissue, it is present at the cell surface at very low levels. In addition, little is known about regulation of ANO1 traffic, namely which factors promote its plasma membrane (PM) localization. Here, we generated a novel cellular model, expressing an inducible 3HA-ANO1-eGFP construct, and validated its usage as a microscopy tool to monitor for ANO1 traffic. We demonstrate the robustness and specificity of this cell-based assay, by the identification of siRNAs acting both as ANO1 traffic enhancer and inhibitor, targeting respectively COPB1 and ESYT1 (extended synaptotagmin-1), the latter involved in coupling of the endoplasmic reticulum to the PM at specific microdomains. We further show that knockdown of ESYT1 (and family members ESYT2 and ESYT3) significantly decreased ANO1 current density. This ANO1 cell-based assay constitutes an important tool to be further used in high-throughput screens and drug discovery of high relevance for CF and cancer.

Date of Submisson/Request
2017-06-12
Date of Acceptance
2017-11-14
Date of Publication
2017-11-15

Institution
BioISI - Biosystems & Integrative Sciences Institute

Where published
Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

Publication Identifiers
ISSN - 0167-4889

Publisher
Elsevier BV

Volume
1865
Number
2

Starting page
421
Last page
431

Document Identifiers
URL - http://dx.doi.org/10.1016/j.bbamcr.2017.11.009
DOI - https://doi.org/10.1016/j.bbamcr.2017.11.009

Rankings
SCIMAGO Q1 (2018) - 2.092 - Cell Biology
SCIMAGO Q1 (2018) - 2.092 - Molecular Biology
Web Of Science Q1 (2018) - 4.739 - BIOCHEMISTRY & MOLECULAR BIOLOGY - SCIE

Keywords
Endoplasmic reticulum Intracellular traffic Calcium-activated chloride channels Cystic fibrosis Automated fluorescence microscopy


Export

APA
Joana R. Lérias, Madalena C. Pinto, Hugo M. Botelho, Nikhil T. Awatade, Margarida C. Quaresma, Iris A.L. Silva, Podchanart Wanitchakool, Rainer Schreiber, Rainer Pepperkok, Karl Kunzelmann, Margarida D. Amaral, (2017). A novel microscopy-based assay identifies extended synaptotagmin-1 (ESYT1) as a positive regulator of anoctamin 1 traffic. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1865, 421-431. ISSN 0167-4889. eISSN . http://dx.doi.org/10.1016/j.bbamcr.2017.11.009

IEEE
Joana R. Lérias, Madalena C. Pinto, Hugo M. Botelho, Nikhil T. Awatade, Margarida C. Quaresma, Iris A.L. Silva, Podchanart Wanitchakool, Rainer Schreiber, Rainer Pepperkok, Karl Kunzelmann, Margarida D. Amaral, "A novel microscopy-based assay identifies extended synaptotagmin-1 (ESYT1) as a positive regulator of anoctamin 1 traffic" in Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, vol. 1865, pp. 421-431, 2017. 10.1016/j.bbamcr.2017.11.009

BIBTEX
@article{37959, author = {Joana R. Lérias and Madalena C. Pinto and Hugo M. Botelho and Nikhil T. Awatade and Margarida C. Quaresma and Iris A.L. Silva and Podchanart Wanitchakool and Rainer Schreiber and Rainer Pepperkok and Karl Kunzelmann and Margarida D. Amaral}, title = {A novel microscopy-based assay identifies extended synaptotagmin-1 (ESYT1) as a positive regulator of anoctamin 1 traffic}, journal = {Biochimica et Biophysica Acta (BBA) - Molecular Cell Research}, year = 2017, pages = {421-431}, volume = 1865 }