PXD021444

PXD021444 is an original dataset announced via ProteomeXchange.

Title	A trimeric Rab7 GEF controls NPC1-dependent lysosomal cholesterol export
Description	Cholesterol import in mammalian cells is mediated by the LDL receptor pathway. Here, we perform a genome-wide CRISPR screen using an endogenous cholesterol reporter and identify >100 genes involved in LDL-cholesterol import. We characterise C18orf8 as a core subunit of the mammalian Mon1-Ccz1 guanidine exchange factor (GEF) for Rab7, required for complex stability and function. C18orf8-deficient cells lack Rab7 activation and show severe defects in late endosome morphology and endosomal LDL trafficking, resulting in cellular cholesterol deficiency. Unexpectedly, free cholesterol accumulates within swollen lysosomes, suggesting a critical defect in lysosomal cholesterol export. We find that active Rab7 interacts with the NPC1 cholesterol transporter and licenses lysosomal cholesterol export. This process is abolished in C18orf8-, Ccz1- and Mon1A/B-deficient cells and restored by a constitutively active Rab7. The trimeric Mon1-Ccz1-C18orf8 (MCC) GEF therefore plays a central role in cellular cholesterol homeostasis coordinating Rab7 activation, endosomal LDL trafficking and NPC1-dependent lysosomal cholesterol export.
HostingRepository	PRIDE
AnnounceDate	2020-10-29
AnnouncementXML	Submission_2020-11-09_04:41:56.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD021444
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	James Williamson
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2020-09-13 23:11:44	ID requested
1	2020-10-29 06:22:13	announced
⏵ 2	2020-11-09 04:41:57	announced	2020-11-09: Updated publication reference for PubMed record(s): 33144569.

van den Boomen DJH, Sienkiewicz A, Berlin I, Jongsma MLM, van Elsland DM, Luzio JP, Neefjes JJC, Lehner PJ, A trimeric Rab7 GEF controls NPC1-dependent lysosomal cholesterol export. Nat Commun, 11(1):5559(2020) [pubmed]

submitter keyword: C18orf8, Mon1, Ccz1, Rab7, GEF, LDL, cholesterol, NPC1, lysosome

Paul Lehner
contact affiliation	Cambridge Institute of Therapeutic Immunology and Infectious Disease, University of Cambridge, UK
contact email	pjl30@cam.ac.uk
lab head
James Williamson
contact affiliation	University of Cambridge
contact email	jcw76@cam.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD021444
     1. Label: PRIDE project
     2. Name: A trimeric Rab7 GEF controls NPC1-dependent lysosomal cholesterol export