Two Saccharomyces cerevisiae strains, BY4741 and BY4741-derived strain lacking the IST2 gene (ist2Δ), were used to characterise the possible role of cortical endoplasmic reticulum (ER) protein Ist2 upon cell dehydration and subsequent rehydration. For the first time, we show that not only protein components of the plasma membrane (PM), but also at least one ER membrane protein (Ist2) play an important role in the maintenance of the viability of yeast cells during dehydration and subsequent rehydration. The low viability of the mutant strain ist2∆ upon dehydration-rehydration stress was related to the lack of Ist2 protein in the ER. We revealed that the PM of ist2∆ strain is not able to completely restore its molecular organisation during reactivation from the state of anhydrobiosis. As the result, the permeability of the PM remains high regardless of the type of reactivation (rapid or gradual rehydration). We conclude that ER protein Ist2 plays an important role in ensuring the stability of molecular organisation and functionality of the PM during dehydration-rehydration stress. These results indicate an important role of ER-PM interactions during cells transition into the state of anhydrobiosis and the subsequent restoration of their physiological activities.

Department of Membrane Transport Institute of Physiology Czech Academy of Sciences Prague Videnska 1083 142 20 Prague 4 Czech Republic

Laboratory of Cell Biology Institute of Microbiology and Biotechnology University of Latvia Jelgavas Str 1 537 Riga 1004 Latvia

Zobrazit více v PubMed

Beker MJ, Rapoport AI (1987) Conservation of yeast by dehydration. Adv Biochem Eng Biotechnol 35:127–171

Beker MJ, Blumbergs JE, Ventina EJ, Rapoport AI (1984) Characteristics of celluar membranes at rehydration of dehydrated yeast Saccharomyces cerevisiae. Appl Microbiol Biotehnol 19:347–352

Bernauer L, Radkohl A, Lehmayer LGK, Emmerstorfer-Augustin A (2021) Komagataella phaffii as emerging model organism in fundamental research. Front Microbiol 11:607028 PubMed PMC

Borovikova D, Herynkova P, Rapoport A, Sychrova H (2014) Potassium uptake system Trk2 is crucial for yeast cell viability during anhydrobiosis. FEMS Microbiol Lett 350:28–33 PubMed

Borovikova D, Teparic R, Mrsa V, Rapoport A (2016) Anhydrobiosis in yeast: cell wall mannoproteins are important for yeast Saccharomyces cerevisiae resistance to dehydration. Yeast 33:347–353 PubMed

Calahan D, Dunham M, DeSevo C, Koshland DE (2011) Genetic analysis of desiccation tolerance in Saccharomyces cerevisiae. Genetics 189:507–519 PubMed PMC

Camara AA Jr, Sant’ Ana AS (2021) Advances in yeast preservation: physiological aspects for cell perpetuation. Curr Opin Food Sci 38:62–70

Chang CL, Chen YJ, Liou J (2017) ER-plasma membrane junctions: why and how do we study them? BBA Mol Cell Res 1864:1494–1506

Choudhary V, Schneiter R (2020) Lipid droplet biogenesis from specialized ER subdomains. Microbial Cell 7:218–221 PubMed PMC

Choudhary V, El Atab O, Mizzon G, Prinz WA, Schneiter R (2020) Seipin and Nem1 establish discrete ER subdomains to initiate yeast lipid droplet biogenesis. J Cell Biol 219:e201910177 PubMed PMC

Clegg JS (2001) Cryptobiosis–a peculiar state of biological organization. Comp Biochem Physiol Part B 128:613–624

Crowe JH, Crowe LM, Carpenter JF, Wistrom CA (1987) Stabilization of dry phospholipid bilayers and proteins by sugars. Biochem J 242:1–10 PubMed PMC

Crowe JH, Hoekstra FA, Crowe LM (1989) Membrane phase transitions are responsible for imbibitional damage in dry pollen. PNAS 86:520–523 PubMed PMC

Crowe JH, Oliver AE, Tablin F (2002) Is there a single biochemical adaptation to anhydrobiosis. Integr Comp Biol 42:497–503 PubMed

Dupont S, Beney L, Ritt JF, Lherminier J, Gervais P (2010) Lateral reorganization of plasma membrane is involved in the yeast resistance to severe dehydration. Biochim Biophys Acta 1798:975–985 PubMed

Dupont S, Rapoport A, Gervais P, Beney L (2014) The survival kit of Saccharomyces cerevisiae for anhydrobiosis. Appl Microbiol Biotechnol 98:8821–8834 PubMed

Duskova M, Borovikova D, Herynkova P, Rapoport A, Sychrova H (2015) The role of glycerol transporters in yeast cells in various physiological and stress conditions. FEMS Microbiol Lett 362:1–8 PubMed

Fischer MA, Temmerman K, Ercan E, Nickel W, Seedorf M (2009) Binding of plasma membrane lipids recruits the yeast integral membrane protein Ist2 to the cortical ER. Traffic 10:1084–1097 PubMed

Grube M, Gavare M, Rozenfelde L, Rapoport A (2014) Anhydrobiosis in yeast: FT-IR spectroscopic studies of yeast grown under conditions of severe oxygen limitation. Biotechnol Appl Biochem 61:474–479 PubMed

Guzhova I, Krallish I, Khroustalyova G, Margulis B, Rapoport A (2008) Dehydration of yeast: changes in the intracellular content of Hsp 70 family proteins. Proc Biochem 43:1138–1141

Hansen KG, Aviram N, Laborenz J, Bibi C, Meyer M, Spang A, Schuldiner M, Herrmann JM (2018) An ER surface retrieval pathway safeguards the import of mitochondrial membrane proteins in yeast. Science 361:1118–1122 PubMed

Hijazi I, Knupp J, Chang A (2020) Retrograde signaling mediates an adaptive survival response to endoplasmic reticulum stress in Saccharomyces cerevisiae. J Cell Sci 133:jcs241539 PubMed PMC

Kaufman RJ, Malhotra JD (2014) Calcium trafficking integrates endoplasmic reticulum function with mitochondrial bioenergetics. Biochim Biophys Acta 1843:2233–2239 PubMed PMC

Kawazoe N, Kimata Y, Izawa S (2017) Acetic acid causes endoplasmic reticulum stress and induces the unfolded protein response in Saccharomyces cerevisiae. Front Microbiol 8:1192 PubMed PMC

Knupp J, Arvan P, Chang A (2019) Increased mitochondrial respiration promotes survival from endoplasmic reticulum stress. Cell Death Differ 26:487–501 PubMed

Kralt A, Carreta M, Mari M, Reggiori F, Steen A, Poolman B, Veenhoff LM (2014) Intrinsically disordered linker and plasma membrane-binding motif sort Ist2 and Ssy1 to junctions. Traffic 16:135–147 PubMed

Ktistakis NT (2020) ER platforms mediating autophagosome generation. BBA Mol Cell Biol Lipids 1865:158433

Kulikova-Borovikova D, Lisi S, Dauss E, Alamae T, Buzzini P, Hallsworth JE, Rapoport A (2018) Activity of the α-glucoside transporter Agt1 in Saccharomyces cerevisiae cells during dehydration-rehydration events. Fungal Biol 122:613–620 PubMed

Le QG, Ishiwata-Kimata Y, Kohno K, Kimata Y (2016) Cadmium impairs protein folding in the endoplasmic reticulum and induces the unfolded protein response. FEMS Yeast Res 16:049

Leslie SB, Teter SA, Crowe LM, Crowe JH (1994) Trehalose lowers membrane phase transitions in dry yeast cells. Biochim Biophys Acta 1192:7–13 PubMed

Manford AG, Stefan CJ, Yuan HL, MacGrun JA, Emr SD (2012) ER-to-plasma membrane tethering proteins regulate cell signaling and ER morphology. Dev Cell 23:1129–1140 PubMed

Miyagawa K-I, Ishiwata-Kimata Y, Kohno K, Kimata Y (2014) Ethanol stress impairs protein folding in the endoplasmic reticulum and activates Ire1 in Saccharomyces cerevisiae. Biosci Biotechnol Biochem 78:1389–1391 PubMed

Nakatogawa H (2020) Autophagic degradation of the endoplasmic reticulum. Proc Jpn Acad Ser B 96:1–9

Nakatsukasa K (2021) Potential physiological relevance of ERAD to the biosynthesis of GPI-anchored proteins in yeast. Int J Mol Sci 22:1061 PubMed PMC

Navarette C, Petrezsélyová S, Barreto L, Martínez JL, Zahrádka J, Ariño J, Sychrová H, Ramos J (2010) Lack of main K

Nguyen TSL, Kohno K, Kimata Y (2013) Zinc depletion activates the endoplasmic reticulum-stress sensor Ire1 via pleiotropic mechanisms. Biosci Biotechnol Biochem 77:1337–1339 PubMed

Papoušková K, Andrsova M, Sychrová H (2017) Lack of cortical endoplasmic reticulum protein Ist2 alters sodium accumulation in Saccharomyces cerevisiae cells. FEMS Yeast Res 17:fox011

Phillips MJ, Voeltz GK (2016) Structure and function of ER membrane contact sites with other organelles. Nat Rev Mol Cell Biol 17:69–82 PubMed

Rapoport A (2017) Anhydrobiosis and dehydration of yeast. In: Sibirny A (ed) Biotechnology of yeast and filamentous fungi. Springer, New York, pp 87–116

Rapoport A (2019) Anhydrobiosis in non-conventional yeasts. In: Sibirny A (ed) Non-conventional yeasts: from basic research to application. Springer, New York, pp 341–359

Rapoport AI, Meissel MN (1985) Survival rates of yeast organisms after dehydration as determined by fluorescence microscopy. Mikrobiologiya 54:66–68

Rapoport AI, Khrustaleva GM, Chamanis GI, Beker ME (1995) Yeast anhydrobiosis: permeability of the plasma membrane. Mikrobiologiya 64:275–278

Rapoport AI, Khroustalyova GM, Kuklina EN (1997) Anhydrobiosis in yeast: activation effect. Brazil J Med Biol Res 30:9–13

Rapoport AI, Khroustalyova GM, Crowe LM, Crowe JH (2009) Anhydrobiosis in yeast: stabilization by exogenous lactose. Microbiol 78:624–629

Rapoport A, Rusakova A, Khroustalyova G, Walker G (2014) Thermotolerance in Saccharomyces cerevisiae is linked to resistance to anhydrobiosis. Proc Biochem 49:1889–1892

Rapoport A, Turchetti B, Buzzini P (2016) Application of anhydrobiosis and dehydration of yeasts for non-conventional biotechnological goals. World J Microbiol Biotechnol 32:1–10

Rapoport A, Golovina EA, Gervais P, Dupont S, Beney L (2019) Anhydrobiosis: inside yeast cells. Biotechnol Adv 37:51–67 PubMed

Ren Q, Brenner R, Boothby TC, Zhang Z (2020) Membrane and lipid metabolism plays an important role in desiccation resistance in the yeast Saccharomyces cerevisiae. BMC Microbiol 20:338 PubMed PMC

Rozenfelde L, Rapoport A (2014) Anhydrobiosis in yeast: is it possible to reach anhydrobiosis for yeast grown in conditions with severe oxygen limitation? Antonie van Leeuwenhoek J Microbiol 106:211–217 PubMed

Smethurst DGJ, Cooper KF (2017) ER fatalities—the role of ER-mitochondrial contact sites in yeast life and death decisions. Mech Ageing Dev 161:225–233 PubMed

Tyo KEJ, Liu Z, Petranovic D, Nielsen J (2012) Imbalance of heterologous protein folding and disulfide bond formation rates yields runaway oxidative stress. BMC Biol 10:16 PubMed PMC

van Leeuwenhoek A (1705) A letter to the royal society concerning animalcules on the roofs of duck-weed. Philos Trans R Soc Lond 24(296):1784–1793

van Leeuwenhoek A (1702) On certain animalcules found in the sediment in gutters of the roofs of houses. Letter 144 in The Select works of Antony van Leeuwenhoek, translated by Samuel Hoole, 1798, 1807, 2 vols. London. 2: 207–213

Voeltz GK, Rolls MM, Rapoport TA (2002) Structural organization of the endoplasmic reticulum. EMBO Rep 3:944–950 PubMed PMC

Walter P, Ron D (2011) The unfolded protein response: from stress pathway to homeostatic regulation. Science 334:1081–1086 PubMed

West M, Zurek N, Hoenger A, Voeltz GK (2011) A 3D analysis of yeast ER structure reveals how ER domains are organized by membrane curvature. J Cell Biol 193:333–346 PubMed PMC

Wolf W, Kilic A, Schrul B, Lorenz H, Schwappach B, Seedorf M (2012) Yeast Ist2 recruits the endoplasmic reticulum to the plasma membrane and creates a ribosome free membrane microcompartment. PLoS ONE 7:1–13

Wolf W, Meese K, Seedorf M (2014) Ist2 in the yeast cortical endoplasmic reticulum promotes trafficking of the amino acid transporter Bap2 to the plasma membrane. PLoS ONE 9:1–13