7
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: not found

      Multilevel regulation of an α-arrestin by glucose depletion controls hexose transporter endocytosis

      research-article

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Changes in nutrient availability trigger massive rearrangements of the yeast plasma membrane proteome. This work shows that the arrestin-related protein Csr2/Art8 is regulated by glucose signaling at multiple levels, allowing control of hexose transporter ubiquitylation and endocytosis upon glucose depletion.

          Abstract

          Nutrient availability controls the landscape of nutrient transporters present at the plasma membrane, notably by regulating their ubiquitylation and subsequent endocytosis. In yeast, this involves the Nedd4 ubiquitin ligase Rsp5 and arrestin-related trafficking adaptors (ARTs). ARTs are targeted by signaling pathways and warrant that cargo ubiquitylation and endocytosis appropriately respond to nutritional inputs. Here, we show that glucose deprivation regulates the ART protein Csr2/Art8 at multiple levels to trigger high-affinity glucose transporter endocytosis. Csr2 is transcriptionally induced in these conditions through the AMPK orthologue Snf1 and downstream transcriptional repressors. Upon synthesis, Csr2 becomes activated by ubiquitylation. In contrast, glucose replenishment induces CSR2 transcriptional shutdown and switches Csr2 to an inactive, deubiquitylated form. This glucose-induced deubiquitylation of Csr2 correlates with its phospho-dependent association with 14-3-3 proteins and involves protein kinase A. Thus, two glucose signaling pathways converge onto Csr2 to regulate hexose transporter endocytosis by glucose availability. These data illustrate novel mechanisms by which nutrients modulate ART activity and endocytosis.

          Related collections

          Most cited references102

          • Record: found
          • Abstract: found
          • Article: not found

          A generic protein purification method for protein complex characterization and proteome exploration.

          We have developed a generic procedure to purify proteins expressed at their natural level under native conditions using a novel tandem affinity purification (TAP) tag. The TAP tag allows the rapid purification of complexes from a relatively small number of cells without prior knowledge of the complex composition, activity, or function. Combined with mass spectrometry, the TAP strategy allows for the identification of proteins interacting with a given target protein. The TAP method has been tested in yeast but should be applicable to other cells or organisms.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds.

            An expression system for Saccharomyces cerevisiae (Sc) has been developed which, depending on the chosen vector, allows the constitutive expression of proteins at different levels over a range of three orders of magnitude and in different genetic backgrounds. The expression system is comprised of cassettes composed of a weak CYC1 promoter, the ADH promoter or the stronger TEF and GPD promoters, flanked by a cloning array and the CYC1 terminator. The multiple cloning array based on pBIISK (Stratagene) provides six to nine unique restriction sites, which facilitates the cloning of genes and allows for the directed cloning of cDNAs by the widely used ZAP system (Stratagene). Expression cassettes were placed into both the centromeric and 2 mu plasmids of the pRS series [Sikorski and Hieter, Genetics 122 (1989) 19-27; Christianson et al., Gene 110 (1992) 119-122] containing HIS3, TRP1, LEU2 or URA3 markers. The 32 expression vectors created by this strategy provide a powerful tool for the convenient cloning and the controlled expression of genes or cDNAs in nearly every genetic background of the currently used Sc strains.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation.

              Networks of protein interactions coordinate cellular functions. We describe a bimolecular fluorescence complementation (BiFC) assay for determination of the locations of protein interactions in living cells. This approach is based on complementation between two nonfluorescent fragments of the yellow fluorescent protein (YFP) when they are brought together by interactions between proteins fused to each fragment. BiFC analysis was used to investigate interactions among bZIP and Rel family transcription factors. Regions outside the bZIP domains determined the locations of bZIP protein interactions. The subcellular sites of protein interactions were regulated by signaling. Cross-family interactions between bZIP and Rel proteins affected their subcellular localization and modulated transcription activation. These results attest to the general applicability of the BiFC assay for studies of protein interactions.
                Bookmark

                Author and article information

                Journal
                J Cell Biol
                J. Cell Biol
                jcb
                jcb
                The Journal of Cell Biology
                The Rockefeller University Press
                0021-9525
                1540-8140
                05 June 2017
                : 216
                : 6
                : 1811-1831
                Affiliations
                [1 ]Institut Jacques Monod, UMR 7592 Centre National de la Recherche Scientifique/Université Paris-Diderot, Sorbonne Paris Cité, 75013 Paris, France
                [2 ]Proteomics Facility, Institut Jacques Monod, UMR 7592 Centre National de la Recherche Scientifique/Université Paris-Diderot, Sorbonne Paris Cité, 75013 Paris, France
                [3 ]Institute of Microbiology of the Czech Academy of Sciences, v.v.i. BIOCEV, 252 50 Vestec, Czech Republic
                [4 ]Faculty of Science, Charles University, BIOCEV, 252 50 Vestec, Czech Republic
                Author notes
                Correspondence to Sébastien Léon: sebastien.leon@ 123456ijm.fr
                [*]

                Q. Defenouillère and V. Albanèse contributed equally to this paper.

                Author information
                http://orcid.org/0000-0003-2255-4859
                http://orcid.org/0000-0002-5947-7415
                http://orcid.org/0000-0002-2536-8595
                Article
                201610094
                10.1083/jcb.201610094
                5461024
                28468835
                63842be7-6b2f-44de-936c-3bada63ee0c1
                © 2017 Hovsepian et al.

                This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

                History
                : 26 October 2016
                : 03 February 2017
                : 28 March 2017
                Funding
                Funded by: Ministère de l’Enseignement Supérieur et de la Recherche, DOI http://dx.doi.org/10.13039/501100004792;
                Funded by: Fondation ARC pour la Recherche sur le Cancer, DOI http://dx.doi.org/10.13039/501100004097;
                Award ID: DOC20160604208
                Funded by: Fondation pour la Recherche Médicale, DOI http://dx.doi.org/10.13039/501100002915;
                Award ID: SPF20150934065
                Funded by: Fondation ARC pour la Recherche sur le Cancer, DOI http://dx.doi.org/10.13039/501100004097;
                Award ID: PJA20151203368
                Funded by: Ligue Contre le Cancer, DOI http://dx.doi.org/10.13039/501100004099;
                Award ID: RS14/75-120
                Funded by: Agence Nationale pour la Recherche, DOI http://dx.doi.org/10.13039/501100001665;
                Award ID: ANR-16-CE13-0002-01
                Funded by: Czech Ministry of Education Youth and Sports, DOI http://dx.doi.org/10.13039/501100001823;
                Award ID: LQ1604 NPU II
                Funded by: Czech Science Foundation, DOI http://dx.doi.org/10.13039/501100001824;
                Award ID: 15-08225S
                Funded by: EU COST Action
                Award ID: PROTEOSTASIS BM1307
                Categories
                Research Articles
                Article
                34
                29
                44

                Cell biology
                Cell biology

                Comments

                Comment on this article