PXD042710

PXD042710 is an original dataset announced via ProteomeXchange.

Title	Genome-Wide CRISPR/Cas9 Screen shows that inhibiting GET4 is neuroprotective and increases mitochondrial-endoplasmic reticulum contact sites
Description	Organelles form contacts between each other allowing for transfer of molecules and signals. Mitochondria-ER Contact sites (MERCS) are lengths of the Mitochondria and ER membrane which come together but do not fuse. They have been implicated in many diseases, including neurodegenerative, metabolic, and cardiac disease. MERCS have been highly researched, however, a lot is still left to explore. To uncover novel regulators of MERCS we conducted a genome-wide, flow cytometry-based screen using an engineered MERCs reporter cell line. We found 410 genes whose downregulation promotes MERCS and 230 genes whose downregulation decreases MERCS. From these, 29 genes were selected from each population for arrayed screening. 14/29 validated from the higher population and 7/29 from the low population. GET4 and BAG6 were highlighted as the top 2 genes that upon suppression increase MERCS from both the pooled and arrayed screen, and were subjected to further investigation. Loss of GET4 or BAG6 increased mVenus puncta, Proximity Ligation Assay (PLA) puncta and increased the percentage of ER in contact with mitochondria. In addition, GET4 and BAG6 were observed to interact with known MERCS proteins, inositol 1,4,5-trisphosphate receptors (IP3R) and Glucose-regulated protein 75 (GRP75). In addition, we found that loss of GET4 increased Calcium concentration [Ca2+] in the mitochondria and mitochondrial respiration. Finally, we show that loss of GET4 rescues motor ability, neurodegeneration and improves lifespan in a Drosophila model of Alzheimer's Disease (Aβ42Arc). Together, these results suggest that GET4 is involved in decreasing MERCS and that its loss is neuroprotective.
HostingRepository	PRIDE
AnnounceDate	2024-05-24
AnnouncementXML	Submission_2024-05-24_02:47:51.836.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Robin Antrobus
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2023-06-04 07:42:36	ID requested
⏵ 1	2024-05-24 02:47:52	announced

10.1038/s41419-024-06568-y;

Wilson EL, Yu Y, Leal NS, Woodward JA, Patikas N, Morris JL, Field SF, Plumbly W, Paupe V, Chowdhury SR, Antrobus R, Lindop GE, Adia YM, Loh SHY, Prudent J, Martins LM, Metzakopian E, Genome-wide CRISPR/Cas9 screen shows that loss of GET4 increases mitochondria-endoplasmic reticulum contact sites and is neuroprotective. Cell Death Dis, 15(3):203(2024) [pubmed]

submitter keyword: Mitochondrial-ER contact site, neuroprotection, IP-mass spectrometry

Emmanouil Metzakopian
contact affiliation	UK Dementia Research Institute, University of Cambridge, Clifford Albutt building, Cambridge biomedical campus, Cambridge, CB2 0AH, UK
contact email	mano982@gmail.com
lab head
Robin Antrobus
contact affiliation	CIMR medicine
contact email	pra29@cam.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD042710
     1. Label: PRIDE project
     2. Name: Genome-Wide CRISPR/Cas9 Screen shows that inhibiting GET4 is neuroprotective and increases mitochondrial-endoplasmic reticulum contact sites