Dataset Information

GPI glycan analysis of GPI-anchored protein from yeast

ABSTRACT: Quality controlled lipid-based protein sorting into selective ER exit sites

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Miyako Nakano

PROVIDER: PXD032852 | JPOST Repository | Mon Mar 28 00:00:00 BST 2022

REPOSITORIES: jPOST

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 15

Publications

Quality-controlled ceramide-based GPI-anchored protein sorting into selective ER exit sites.

Rodriguez-Gallardo Sofia S Sabido-Bozo Susana S Ikeda Atsuko A Araki Misako M Okazaki Kouta K Nakano Miyako M Aguilera-Romero Auxiliadora A Cortes-Gomez Alejandro A Lopez Sergio S Waga Miho M Nakano Akihiko A Kurokawa Kazuo K Muñiz Manuel M Funato Kouichi K

Cell reports 20220501 5

Glycosylphosphatidylinositol-anchored proteins (GPI-APs) exit the endoplasmic reticulum (ER) through a specialized export pathway in the yeast Saccharomyces cerevisiae. We have recently shown that a very-long acyl chain (C26) ceramide present in the ER membrane drives clustering and sorting of GPI-APs into selective ER exit sites (ERES). Now, we show that this lipid-based ER sorting also involves the C26 ceramide as a lipid moiety of GPI-APs, which is incorporated into the GPI anchor through a l ...[more]

PMID: 35508142

Similar Datasets

Project description:Maintenance of protein homeostasis is essential for cellular survival. Central to this regulation are mechanisms of protein quality control in which misfolded proteins are recognized and degraded by the ubiquitin-proteasome system. One well-studied protein quality control pathway requires ER resident, multi-subunit E3 ubiquitin ligases that function in ER-associated degradation (ERAD). Using fission yeast, our lab identified the Golgi Dsc E3 ligase as required for proteolytic activation of fungal sterol regulatory element-binding protein (SREBP) transcription factors. The Dsc E3 ligase contains 5 integral membrane subunits and structurally resembles ERAD E3 ligases. S. cerevisiae codes for homologs of Dsc E3 ligase subunits, including the Dsc1 E3 ligase homolog Tul1 that functions in Golgi protein quality control. Interestingly, S. cerevisiae lacks SREBP homologs, indicating that novel Tul1 E3 ligase substrates exist. Here, we show that the S. cerevisiae Tul1 E3 ligase consists of Tul1, Dsc2, Dsc3, and Ubx3 and define Tul1 complex architecture. Tul1 E3 ligase function required each subunit as judged by vacuolar sorting of the artificial substrate Pep12D. Genetic studies demonstrated that Tul1 E3 ligase is required in cells lacking the multivesicular body pathway and under conditions of ubiquitin depletion. To identify candidate substrates, we used quantitative diGly proteomics to survey ubiquitylation in wild-type and tul1∆ cells. We identified 3,116 non-redundant ubiquitylation sites, including 10 sites in candidate substrates. Quantitative proteomics found 4.5% of quantified proteins (53/1172) to be differentially expressed in tul1∆ cells. Correcting the diGly dataset for these differences increased the number of Tul1-dependent ubiquitylation sites. Together, these data demonstrate that the Tul1 E3 ligase plays a minor role in protein homeostasis under non-stress conditions, consistent with functions in protein quality control. Furthermore, this quantitative diGly proteomics methodology serves as a robust platform to screen for stress conditions that require Tul1 activity.

			Action	DRS
	19031102_GW1.msf	Msf
	19031102_GW1_WT.raw	Raw
	19031103_GtM_ted1_mature.raw	Raw
	19031103_Gtm.msf	Msf
	19031104_GtE.msf	Msf