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LYSET/TMEM251 KO secretome from MIA PaCa-2 cells

ABSTRACT: Analysis of proteins secreted over 24 h in OptiMEM by LYSET/TMEM251 knockout and control MIA PaCa-2 cells.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Pancreas

DISEASE(S): Pancreas Adenocarcinoma

SUBMITTER: Wilhelm Palm

LAB HEAD: Wilhelm Palm

PROVIDER: PXD029658 | Pride | 2022-09-29

REPOSITORIES: Pride

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			Action	DRS
	CP0480-1.zip	Other
	CP480_RawFile2Sample.txt	Txt
	checksum.txt	Txt
	oecf3-CP0480-01.raw	Raw
	oecf3-CP0480-02.raw	Raw

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LYSET/TMEM251 KO cellular and lysosomal proteomes from MIA PaCa-2 cells

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GCAF(TMEM251) regulates lysosome biogenesis by activating the mannose-6-phosphate pathway

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GOLPH3 and GOLPH3L maintain Golgi localization of LYSET and a functional mannose 6-phosphate transport pathway

Project description:Glycosylation plays an important role for modifications of lipids and sorting of proteins that is regulated by asymmetric intra-Golgi distribution and SPPL3-mediated cleavage of Golgi enzymes. We found that cells lacking the Golgi N-acetylglucosamine phosphotransferase (GNPT) retention factor LYSET/TMEM251 display a SPPL3-dependent hypersecretion of the galactosyltransferase B4GALT5 which is caused by the loss of its lysosomal mannose 6-phosphate (M6P) targeting signal. Similarly, loss of the COPI adaptors GOLPH3/GOLPH3L destabilized LYSET-GNPT complexes and led to a prominent hypersecretion of B4GALT5 and lysosomal enzymes. Mechanistically, we identified LYSET as a novel, atypical client of GOLPH3/GOLPH3L. GOLPH3/GOLPH3L hence ensure cis-Golgi localization of the LYSET-GNPT complex and maintain Golgi polarity, integrity of the M6P tagging machinery and homeostasis of lysosomes.

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mRNA expression data from RalGAPβ-deficient human pancreatic ductal adenocarcinoma cells (MIA PaCa-2), MIA PaCa-2 parental cells, and MIA PaCa-2 control cells

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The Human Disease Gene LYSET is Essential for Lysosomal Enzyme Transport and Viral Infection

Project description:Lysosomes are key degradative compartments of the cell. Transport to lysosomes ismediated by tagging soluble enzymes with mannose 6-phosphate (M6P) by GlcNAc-1-phosphotransferase whose deficiency leads to the severe lysosomal storage disorder mucolipidosisII (MLII). Several viruses require lysosomal cathepsins to cleave structural proteins and thusdepend on functional GlcNAc-1-phosphotransferase. Using genome-scale CRISPR screens, weidentify LYSET as essential for infection by cathepsin-dependent viruses including SARS-CoV-2. We show that LYSET deficiency results in global loss of M6P tagging and mislocalization ofGlcNAc-1-phosphotransferase to lysosomes. Lyset knockout mice exhibit MLII-like phenotypesand human pathogenic LYSET alleles fail to restore lysosomal sorting defects. Thus, we uncoverLYSET as an indispensable component of the M6P trafficking machinery and reveal thebiochemical basis of an inherited disease caused by mutations in LYSET.

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| S-EPMC9740945 | biostudies-literature