Intracellular trehalose improves osmotolerance but not desiccation tolerance in mammalian cells

Castro, Arcadio García de; Tunnacliffe, Alan

doi:10.1016/s0014-5793(00)02340-1

Cited by 74 publications

(26 citation statements)

References 31 publications

(31 reference statements)

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“…Protection against more extreme water loss imposed by a Ϸ1,000 mOsm upshift or by desiccation might result from extending LEA protein expression to other compartments in the cell, or by combination with, for example, the capacity to synthesize trehalose (36). Thus, our results offer insight into one of the protective mechanisms in anhydrobiosis, namely protein stabilization, but also highlight a potentially important component for anhydrobiotic engineering of desiccationsensitive cell types to full desiccation tolerance (37).…”

Section: Discussionmentioning

confidence: 77%

Hydrophilic protein associated with desiccation tolerance exhibits broad protein stabilization function

Chakrabortee

Boschetti

Walton

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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273

248

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The ability of certain plants, invertebrates, and microorganisms to survive almost complete loss of water has long been recognized, but the molecular mechanisms of this phenomenon remain to be defined. One phylogenetically widespread adaptation is the presence of abundant, highly hydrophilic proteins in desiccation-tolerant organisms. The best characterized of these polypeptides are the late embryogenesis abundant (LEA) proteins, first described in plant seeds >20 years ago but recently identified in invertebrates and bacteria. The function of these largely unstructured proteins has been unclear, but we now show that a group 3 LEA protein from the desiccation-tolerant nematode Aphelenchus avenae is able to prevent aggregation of a wide range of other proteins both in vitro and in vivo. The presence of water is essential for maintenance of the structure of many proteins, and therefore desiccation stress induces unfolding and aggregation. The nematode LEA protein is able to abrogate desiccation-induced aggregation of the water-soluble proteomes from nematodes and mammalian cells and affords protection during both dehydration and rehydration. Furthermore, when coexpressed in a human cell line, the LEA protein reduces the propensity of polyglutamine and polyalanine expansion proteins associated with neurodegenerative diseases to form aggregates, demonstrating in vivo function of an LEA protein as an antiaggregant. Finally, human cells expressing LEA protein exhibit increased survival of dehydration imposed by osmotic upshift, consistent with a broad protein stabilization function of LEA proteins under conditions of water stress.anhydrobiosis ͉ late embryogenesis abundant protein W ater is essential for life, but some organisms survive desiccation and the dry state for long periods during which metabolism and life processes come to a halt, but resume on rehydration. Desiccation tolerance, or anhydrobiosis (''life without water''), is found across all biological kingdoms, including animals and plants such as the nematode Aphelenchus avenae and the resurrection plant Craterostigma plantagineum (1-3). Investigations into the molecular mechanisms of desiccation tolerance have highlighted the importance of various hydrophilic proteins, chief among which are the late embryogenesis abundant (LEA) proteins (4).LEA proteins have been known for many years to accumulate in maturing plant seeds as they acquire desiccation tolerance (5, 6), but their discovery in invertebrates (7-13) suggests that similar mechanisms govern anhydrobiosis in both animals and plants. LEA proteins are known to be largely unstructured in solution, probably because their extreme hydrophilicity favors association with water over intrachain interactions, but they can show increased folding when dried or when associated with phospholipid bilayers (14-16). Although LEA proteins are widely held to protect cells against water stress, their precise role has been a puzzle since they were first described. Recently, evidence supporting possible functions has bee...

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Section: Discussionmentioning

confidence: 77%

Hydrophilic protein associated with desiccation tolerance exhibits broad protein stabilization function

Chakrabortee

Boschetti

Walton

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

273

248

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“…Freeze-dried human and mammalian platelets (apyrene) could recover after rehydration only when they were loaded intracellularly with a relatively high concentration of trehalose (20 mM) before freeze-drying (Wolkers et al, 2001(Wolkers et al, , 2002. On the other hand, mouse cells containing 10% trehalose, through expressing trehalose phosphate synthase (TPS) intracellularly, cannot survive complete desiccation, although they have increased tolerance of high osmolarity (Garcia de Castro and Tunnacliffe, 2000). It is not still determined whether the importance of intracellular trehalose reported in vertebrate cells is applicable in cells, tissues and individuals of invertebrates at the anhydrobiotic state.…”

Section: Mechanism Of Induction Of Anhy-drobiosismentioning

confidence: 99%

Anhydrobiosis in invertebrates

Watanabe

2006

Appl. Entomol. Zool.

178

158

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Recent work on anhydrobiosis in invertebrates is reviewed. I introduce definition and classification of cryptobiosis, and review the distinctive features and extremely high stress tolerance of anhydrobiotic invertebrates. Most anhydrobiotic invertebrates have evolved various kinds of behavioral, morphological, physiological and physical adaptations to reduce water loss during induction of anhydrobiosis. Trehalose is known as a common compatible solute in anhydrobiotic organisms from unicellular organisms to invertebrates and higher plants. Trehalose may provide effective protection against desiccation because it has superior biochemical and physicochemical properties for stabilizing membranes and biomolecules including proteins and lipids. Recent work also indicates several possible kinds of molecules involved in induction of anhydrobiosis. The adaptations necessary for successful induction of and recovery from anhydrobiosis vary greatly among taxa of invertebrates. Understanding the diversity of anhydrobiosis in invertebrates would be a key to elucidate evolutionary scenarios in anhydrobiosis.

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“…Using this value will likely result in a lower calculated intracellular trehalose concentration than the actual concentration because it includes the volume of osmotically active interorganelle solvents that may or may not be available to the cytosolic trehalose. However, using the osmotically active isotonic volume is an improvement on past studies that used the overall isotonic volume when calculating intracellular trehalose concentrations (Garcia de Castro and Tunnacliffe, 2000;Guo et al, 2000;Puhlev et al, 2001). …”

Section: Calculation Of Intracellular Trehalose Concentrationsmentioning

confidence: 99%

“…Among the key impediments to using trehalose during the desiccation of mammalian cells has been the impermeability of the plasma membrane to this molecule. A number of approaches have been used to load trehalose into mammalian cells to improve desiccation tolerance, including the genetic expression of trehalose synthase genes (Garcia de Castro and Tunnacliffe, 2000;Guo et al, 2000;Puhlev et al, 2001), thermotropic phase transitions (Beattie et al, 1997;Wolkers et al, 2001), osmotic shock (Puhlev et al, 2001), thermal shock (Puhlev et al, 2001), and microinjection (Eroglu et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

“…Another method for the separation and quantification of various saccharides is to analyze the silyl derivatives using gas chromatography (Churms, 1990;Knapp, 1979;Molnár-Perl and Horváth, 1997). Although the extraction of intracellular sugars and the derivitization reactions required to enhance the separation and detection characteristics of saccharides can be time-consuming, efforts to simplify these procedures (Churms, 1990;Molnár-Perl and Horváth, 1997) have resulted in procedures for the quantitative measure of intracellular saccharides (Garcia de Castro and Tunnacliffe, 2000).…”

Section: Introductionmentioning

confidence: 99%

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Measurement of trehalose loading of mammalian cells porated with a metal‐actuated switchable pore

Acker

Young

et al. 2003

Biotech & Bioengineering

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Efforts to improve the tolerance of mammalian cells to desiccation have focused on the role that sugars have in protecting cells from lethal injury. Among the key determinants of desiccation tolerance is the intracellular trehalose concentration, and thus quantifying the amount and rate of trehalose accumulation has now become very critical to the success of these desiccation approaches. We introduced trehalose into 3T3 fibroblasts, human keratinocytes, and rat hepatocytes using a genetically engineered mutant of the pore-forming alpha-hemolysin from Staphylococcus aureus. Manipulating the extracellular Zn(2+) concentration selectively opens and closes this pore ( approximately 2 nm) and enables controlled loading of cells with sugars. We quantified intracellular trehalose using gas chromatography-mass spectroscopy (GC-MS) to examine the trimethylsilyl derivative of intracellular trehalose. Using the GC-MS method, we demonstrate that the switchable characteristics of H5 alpha-hemolysin permit controlled loading of the high concentrations of trehalose (up to 0.5 M) necessary for engineering desiccation tolerance in mammalian cells.

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Intracellular trehalose improves osmotolerance but not desiccation tolerance in mammalian cells

Cited by 74 publications

References 31 publications

Hydrophilic protein associated with desiccation tolerance exhibits broad protein stabilization function

Hydrophilic protein associated with desiccation tolerance exhibits broad protein stabilization function

Anhydrobiosis in invertebrates

Measurement of trehalose loading of mammalian cells porated with a metal‐actuated switchable pore

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