Vacuolar Sorting Determinants: Isolation and Study

2018 ◽  
pp. 21-31
Author(s):  
Bruno Peixoto ◽  
Susana Pereira ◽  
Cláudia Pereira ◽  
José Pissarra
Keyword(s):  
Vacuolar Sorting

scholarly journals Actin and Microtubules Differently Contribute to Vacuolar Targeting Specificity during the Export from the ER

Membranes ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 299
Author(s):  
Monica De Caroli ◽  
Fabrizio Barozzi ◽  
Luciana Renna ◽  
Gabriella Piro ◽  
Gian-Pietro Di Sansebastiano
Keyword(s):  
Spatial Targeting ◽  
Traffic Regulation ◽  
Emergent Features ◽  
Vacuolar Sorting ◽  
Er Export ◽  
Golgi Network ◽  
Relationship Of ◽  
The Relationship ◽  
Fine Tune

Plants rely on both actin and microtubule cytoskeletons to fine-tune sorting and spatial targeting of membranes during cell growth and stress adaptation. Considerable advances have been made in recent years in the comprehension of the relationship between the trans-Golgi network/early endosome (TGN/EE) and cytoskeletons, but studies have mainly focused on the transport to and from the plasma membrane. We address here the relationship of the cytoskeleton with different endoplasmic reticulum (ER) export mechanisms toward vacuoles. These emergent features of the plant endomembrane traffic are explored with an in vivo approach, providing clues on the traffic regulation at different levels beyond known proteins’ functions and interactions. We show how traffic of vacuolar markers, characterized by different vacuolar sorting determinants, diverges at the export from the ER, clearly involving different components of the cytoskeleton.

scholarly journals Anin vivoexpression system for the identification of cargo proteins of vacuolar sorting receptors in Arabidopsis culture cells

2013 ◽  
Vol 75 (6) ◽  
pp. 1003-1017 ◽  
Author(s):  
Jinbo Shen ◽  
Pui Kit Suen ◽  
Xiangfeng Wang ◽  
Youshun Lin ◽  
Sze Wan Lo ◽  
...  
Keyword(s):  
Culture Cells ◽  
Vacuolar Sorting Receptors ◽  
Cargo Proteins ◽  
Vacuolar Sorting

scholarly journals Different sensitivity to wortmannin of two vacuolar sorting signals indicates the presence of distinct sorting machineries in tobacco cells.

1995 ◽  
Vol 130 (6) ◽  
pp. 1307-1318 ◽  
Author(s):  
K Matsuoka ◽  
D C Bassham ◽  
N V Raikhel ◽  
K Nakamura
Keyword(s):  
Secretory Pathway ◽  
Plant Cells ◽  
Specific Inhibitor ◽  
Matrix Proteins ◽  
Wild Type ◽  
Dose Dependency ◽  
Tobacco Cells ◽  
Sorting Signals ◽  
Vacuolar Sorting ◽  
Mature Part

Vacuolar matrix proteins in plant cells are sorted from the secretory pathway to the vacuoles at the Golgi apparatus. Previously, we reported that the NH2-terminal propeptide (NTPP) of the sporamin precursor and the COOH-terminal propeptide (CTPP) of the barley lectin precursor contain information for vacuolar sorting. To analyze whether these propeptides are interchangeable, we expressed constructs consisting of wild-type or mutated NTPP with the mature part of barley lectin and sporamin with CTPP and mutated NTPP in tobacco BY-2 cells. The vacuolar localization of these constructs indicated that the signals were interchangeable. We next analyzed the effect of wortmannin, a specific inhibitor of mammalian phosphatidylinositol (PI) 3-kinase on vacuolar delivery by NTPP and CTPP in tobacco cells. Pulse-chase analysis indicated that 33 microM wortmannin caused almost complete inhibition of CTPP-mediated transport to the vacuoles, while NTPP-mediated transport displayed almost no sensitivity to wortmannin at this concentration. This indicates that there are at least two different mechanisms for vacuolar sorting in tobacco cells, and the CTPP-mediated pathway is sensitive to wortmannin. We compared the dose dependencies of wortmannin on the inhibition of CTPP-mediated vacuolar delivery of proteins and on the inhibition of the synthesis of phospholipids in tobacco cells. Wortmannin inhibited PI 3- and PI 4-kinase activities and phospholipid synthesis. Missorting caused by wortmannin displays a dose dependency that is similar to the dose dependency for the inhibition of synthesis of PI 4-phosphate and major phospholipids. This is different, however, than the inhibition of synthesis of PI 3-phosphate. Thus, the synthesis of phospholipids could be involved in CTPP-mediated vacuolar transport.

Saccharomyces cerevisiae Apl2p, a homologue of the mammalian clathrin AP beta subunit, plays a role in clathrin-dependent Golgi functions

1995 ◽  
Vol 108 (4) ◽  
pp. 1605-1615 ◽  
Author(s):  
M.R. Rad ◽  
H.L. Phan ◽  
L. Kirchrath ◽  
P.K. Tan ◽  
T. Kirchhausen ◽  
...  
Keyword(s):  
Chain Gene ◽  
Beta Subunits ◽  
Temperature Sensitive ◽  
Permissive Temperature ◽  
Intracellular Protein ◽  
Protein Traffic ◽  
Specific Effects ◽  
Yeast Chromosome ◽  
Vacuolar Sorting ◽  
Golgi Network

Clathrin-coated vesicles mediate selective intracellular protein traffic from the plasma membrane and the trans-Golgi network. At these sites, clathrin-associated protein (AP) complexes have been implicated in both clathrin coat assembly and collection of cargo into nascent vesicles. We have found a gene on yeast chromosome XI that encodes a homologue of the mammalian AP beta subunits. Disruptions of this gene, APl2, and a previously identified beta homologue, APl1, have been engineered in cells expressing wild-type (CHC1) or temperature sensitive (chc1-ts) alleles of the clathrin heavy chain gene. APl1 or APl2 disruptions (apl1 delta or apl2 delta) yield no discernable phenotypes in CHC1 strains, indicating that the Apl proteins are not essential for clathrin function. However, the apl2 delta, but not the apl1 delta, allele enhances the growth and alpha-factor pheromone maturation defects of chc1-ts cells. Disruption of APl2 also partially suppresses the vacuolar sorting defect that occurs in chc1-ts cells immediately after imposition of the non-permissive temperature. These Golgi-specific effects of apl2 delta in chc1-ts cells provide evidence that Apl2p is a component of an AP complex that interacts with clathrin at the Golgi apparatus.

scholarly journals N-Linked Glycosylation Modulates Golgi-Independent Vacuolar Sorting Mediated by the Plant Specific Insert

Plants ◽  
2019 ◽  
Vol 8 (9) ◽  
pp. 312 ◽  
Author(s):  
Vanessa Vieira ◽  
Bruno Peixoto ◽  
Mónica Costa ◽  
Susana Pereira ◽  
José Pissarra ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Site Directed Mutagenesis ◽  
Research Gaps ◽  
Glycosylation Pattern ◽  
High Sequence Similarity ◽  
Post Translational Modifications ◽  
Vacuolar Sorting ◽  
Prevacuolar Compartment ◽  
Cardosin A ◽  
Comprehensive Study

In plant cells, the conventional route to the vacuole involves the endoplasmic reticulum, the Golgi and the prevacuolar compartment. However, over the years, unconventional sorting to the vacuole, bypassing the Golgi, has been described, which is the case of the Plant-Specific Insert (PSI) of the aspartic proteinase cardosin A. Interestingly, this Golgi-bypass ability is not a characteristic shared by all PSIs, since two related PSIs showed to have different sensitivity to ER-to-Golgi blockage. Given the high sequence similarity between the PSI domains, we sought to depict the differences in terms of post-translational modifications. In fact, one feature that draws our attention is that one is N-glycosylated and the other one is not. Using site-directed mutagenesis to obtain mutated versions of the two PSIs, with and without the glycosylation motif, we observed that altering the glycosylation pattern interferes with the trafficking of the protein as the non-glycosylated PSI-B, unlike its native glycosylated form, is able to bypass ER-to-Golgi blockage and accumulate in the vacuole. This is also true when the PSI domain is analyzed in the context of the full-length cardosin. Regardless of opening exciting research gaps, the results obtained so far need a more comprehensive study of the mechanisms behind this unconventional direct sorting to the vacuole.

A Vacuolar Sorting Domain May Also Influence the Way in Which Proteins Leave the Endoplasmic Reticulum

2001 ◽  
Vol 13 (9) ◽  
pp. 2021 ◽  
Author(s):  
Kirsi Tormakangas ◽  
Jane L. Hadlington ◽  
Peter Pimpl ◽  
Stefan Hillmer ◽  
Federica Brandizzi ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Vacuolar Sorting ◽  
The Way

scholarly journals Demonstration in Yeast of the Function of BP-80, a Putative Plant Vacuolar Sorting Receptor

2001 ◽  
Vol 13 (4) ◽  
pp. 781-792 ◽  
Author(s):  
David Humair ◽  
Doramys Hernández Felipe ◽  
Jean-Marc Neuhaus ◽  
Nadine Paris
Keyword(s):  
Vacuolar Sorting Receptor ◽  
Vacuolar Sorting ◽  
Sorting Receptor

scholarly journals Arabidopsis TOL Proteins Act as Gatekeepers for Vacuolar Sorting of PIN2 Plasma Membrane Protein

2013 ◽  
Vol 23 (24) ◽  
pp. 2500-2505 ◽  
Author(s):  
Barbara Korbei ◽  
Jeanette Moulinier-Anzola ◽  
Lucinda De-Araujo ◽  
Doris Lucyshyn ◽  
Katarzyna Retzer ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Plasma Membrane Protein ◽  
Vacuolar Sorting

scholarly journals The Yeast Casein Kinase Yck3p Is Palmitoylated, then Sorted to the Vacuolar Membrane with AP-3-dependent Recognition of a YXXϕ Adaptin Sorting Signal

2004 ◽  
Vol 15 (3) ◽  
pp. 1397-1406 ◽  
Author(s):  
Beimeng Sun ◽  
Linyi Chen ◽  
Wei Cao ◽  
Amy F. Roth ◽  
Nicholas G. Davis
Keyword(s):  
Plasma Membrane ◽  
Mammalian Cells ◽  
Casein Kinase ◽  
Vacuolar Membrane ◽  
Sorting Signal ◽  
Casein Kinases ◽  
In Trans ◽  
Vacuolar Sorting ◽  
Cis And Trans ◽  
In Cis

Our previous work found the two yeast plasma membrane-localized casein kinases Yck1p and Yck2p to be palmitoylated on C-terminal Cys-Cys sequences by the palmitoyl transferase Akr1p. The present work examines a third casein kinase, Yck3p, which ends with the C-terminal sequence Cys-Cys-Cys-Cys-Phe-Cys-Cys-Cys. Yck3p is palmitoylated and localized to the vacuolar membrane. While the C-terminal cysteines are required for this palmitoylation, Akr1p is not. Palmitoylation requires the C-terminal Yck3p residues 463-524, whereas information for vacuolar sorting maps to the 409-462 interval. Vacuolar sorting is disrupted in cis through deletion of the 409-462 sequences and in trans through mutation of the AP-3 adaptin complex; both cis- and trans-mutations result in Yck3p missorting to the plasma membrane. This missorted Yck3p restores 37°C viability to yck1Δ yck2-ts cells. yck1Δ yck2-ts suppressor mutations isolated within the YCK3 gene identify the Yck3p vacuolar sorting signal—the tetrapeptide YDSI, a perfect fit to the YXXϕ adaptin-binding consensus. Although YXXϕ signals have a well-appreciated role in the adaptin-mediated sorting of mammalian cells, this is the first signal of this class to be identified in yeast.

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