New factors for protein transport identified by a genome-wide CRISPRi screen in mammalian cells

Laia Bassaganyas; Stephanie J Popa; Max Horlbeck; Claudia Puri; Sarah E Stewart; Felix Campelo; Anupama Ashok; Cristian M Butnaru; Nathalie Brouwers; Kartoosh Heydari; Jean Ripoche; Jonathan Weissman; David C Rubinsztein; Randy Schekman; Vivek Malhotra; Kevin Moreau; Julien Villeneuve

doi:10.1083/jcb.201902028

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New factors for protein transport identified by a genome-wide CRISPRi screen in mammalian cells

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New factors for protein transport identified by a genome-wide CRISPRi screen in mammalian cells

Laia Bassaganyas, Stephanie J Popa, Max Horlbeck, Claudia Puri, Sarah E Stewart, Felix Campelo, Anupama Ashok, Cristian M Butnaru, Nathalie Brouwers, Kartoosh Heydari, …

The Journal of cell biology, Vol.218(11), pp.3861-3879

04/11/2019

DOI: https://doi.org/10.1083/jcb.201902028

PMID: 31488582

Résumé

Carrier Proteins - genetics

Carrier Proteins - metabolism

Cell Cycle Proteins - genetics

Cell Cycle Proteins - metabolism

Cells, Cultured

Clustered Regularly Interspaced Short Palindromic Repeats - genetics

Golgi Apparatus - metabolism

HEK293 Cells

HeLa Cells

Humans

Protein and membrane trafficking pathways are critical for cell and tissue homeostasis. Traditional genetic and biochemical approaches have shed light on basic principles underlying these processes. However, the list of factors required for secretory pathway function remains incomplete, and mechanisms involved in their adaptation poorly understood. Here, we present a powerful strategy based on a pooled genome-wide CRISPRi screen that allowed the identification of new factors involved in protein transport. Two newly identified factors, TTC17 and CCDC157, localized along the secretory pathway and were found to interact with resident proteins of ER-Golgi membranes. In addition, we uncovered that upon TTC17 knockdown, the polarized organization of Golgi cisternae was altered, creating glycosylation defects, and that CCDC157 is an important factor for the fusion of transport carriers to Golgi membranes. In conclusion, our work identified and characterized new actors in the mechanisms of protein transport and secretion and opens stimulating perspectives for the use of our platform in physiological and pathological contexts.

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Détails

Titre: New factors for protein transport identified by a genome-wide CRISPRi screen in mammalian cells
Créateurs - sans rôle: Laia Bassaganyas - Department of Medical Genetics, National Institute for Health Research Cambridge Biomedical Research Centre, and Cancer Research UK Cambridge Centre, University of Cambridge, Cambridge, UK
Stephanie J Popa - University of Cambridge Metabolic Research Laboratories, Wellcome Trust-Medical Research Council Institute of Metabolic Science, University of Cambridge, Cambridge, UK
Max Horlbeck - Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA
Claudia Puri - Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK
Sarah E Stewart - University of Cambridge Metabolic Research Laboratories, Wellcome Trust-Medical Research Council Institute of Metabolic Science, University of Cambridge, Cambridge, UK
Felix Campelo - Institut de Ciencies Fotoniques, Barcelona Institute of Science and Technology, Castelldefels, Spain
Anupama Ashok - Centre for Genomic Regulation, Barcelona Institute of Science and Technology, Barcelona, Spain
Cristian M Butnaru - Department of Photonic Investigations, Center of Advanced Laser Technologies, National Institute for Laser, Plasma and Radiation Physics, Magurele, Romania
Nathalie Brouwers - Centre for Genomic Regulation, Barcelona Institute of Science and Technology, Barcelona, Spain
Kartoosh Heydari - Cell Therapy Facility, Stanford Health Care, Stanford, CA
Jean Ripoche - Institut National de la Sante et de la Recherche Medicale U1026, Université de Bordeaux, Bordeaux, France
Jonathan Weissman - Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA
David C Rubinsztein - UK Dementia Research Institute
Randy Schekman - Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA
Vivek Malhotra - Centre for Genomic Regulation, Barcelona Institute of Science and Technology, Barcelona, Spain
Kevin Moreau - University of Cambridge Metabolic Research Laboratories, Wellcome Trust-Medical Research Council Institute of Metabolic Science, University of Cambridge, Cambridge, UK
Julien Villeneuve - Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK
Détails de publication: The Journal of cell biology, Vol.218(11), pp.3861-3879
Note de subvention: Wellcome Trust BB/P010911/1 / Biotechnology and Biological Sciences Research Council MC_UU_12012/5 / Medical Research Council T32 GM007618 / NIGMS NIH HHS T32 EB009383 / NIBIB NIH HHS 109152/Z/15/Z / Wellcome Trust UKDRI-2002 / Medical Research Council
Identifiants: 99162980609311
Unité académique: Institut de Génomique Fonctionnelle - IGF
Langue: English
Type de ressource: Journal article