Evidence for Protein Damage at Environmental Temperatures: Seasonal Changes in Levels of Ubiquitin Conjugates and Hsp70 in the Intertidal Mussel Mytilus Trossulus

Journal of Experimental Biology - Tập 198 Số 7 - Trang 1509-1518 - 1995
Gretchen E. Hofmann1, George N. Somero1
1Oregon State University Department of Zoology , , Corvallis, Oregon 97331-2914, USA

Tóm tắt

ABSTRACT We examined the seasonal variation in environmentally induced protein damage in natural populations of the intertidal mussel Mytilus trossulus. In order to compare the state of protein pools during seasonal variations in environmental temperature, we used solid-phase immunochemical analysis to quantify ubiquitin conjugate concentrations and relative levels of the stress protein hsp70. The two biochemical indices were selected for their cellular roles in irreversible and reversible protein denaturation, respectively. Proteins that are ubiquitinated are irreversibly damaged and are degraded by intracellular proteases; stress proteins act as molecular chaperones to re-fold thermally denatured proteins and, thus, indicate degrees of reversible protein damage. Comparisons involved mussels collected in February and August from two study sites: an intertidal site which subjected animals to a wide range of body temperatures (from approximately 10 to 35 °C in summer), and a subtidal site where animals remained submerged throughout the tidal cycle. Our results show that quantities of ubiquitin conjugates and hsp70 were greater in gill tissue from summer-collected mussels than in gills of winter-collected specimens. Ubiquitin conjugate and hsp70 levels were also greater in mussels collected from an intertidal location than in mussels from a submerged population. Our results show that the high summer temperatures normally experienced in the field are sufficient to cause increased denaturation of cellular proteins. Despite increases in the concentrations of heat shock proteins in summer-acclimatized mussels, elevated levels of irreversibly denatured, i.e. ubiquitinated, proteins were still observed, which indicates that the heat shock response may not be able to rescue all heat-damaged proteins. The energy costs associated with replacing heat-damaged proteins and with maintaining the concentrations and activities of heat shock proteins may contribute substantially to cellular energy demands. These increased energy demands may have an impact on the ecological energetic relationships of species, e.g. in the allocations of energy for growth and reproduction, and, as a consequence, may contribute to determining their distribution limits.

Từ khóa


Tài liệu tham khảo

Ananthan, 1986, Abnormal proteins serve as eukaryotic stress signals and trigger the activation of heat shock genes, Science, 232, 522, 10.1126/science.3083508

Anchordoguy, 1994, Acute blockage of the ubiquitin-mediated proteolytic pathway during invertebrate quiescence, Am. J. Physiol., 267, R895

Bayne, 1976, The physiological ecology of Mytilus calfornianus Conrad. II. Adaptations to low oxygen tension and air exposure, Oecologia, 22, 229, 10.1007/BF00344794

Becker, 1994, Heat-shock proteins as molecular chaperones, Eur. J. Biochem., 219, 11, 10.1111/j.1432-1033.1994.tb19910.x

Beckmann, 1990, Interaction of hsp70 with newly synthesized proteins: implications for protein folding and assembly, Science, 248, 850, 10.1126/science.2188360

Beers, 1993, Utility of polyhistidine-tagged ubiquitin in the purification of ubiquitin–protein conjugates and as an affinity ligand for the purification of ubiquitin-specific hydrolases, J. biol. Chem., 268, 21645, 10.1016/S0021-9258(20)80590-0

Beynon, 1986, Catabolism of intracellular proteins: molecular aspects, Am. J. Physiol., 251, C141, 10.1152/ajpcell.1986.251.2.C141

Bond, 1988, Ubiquitin in stressed chicken embryo fibroblasts, J. biol. Chem., 263, 2384, 10.1016/S0021-9258(18)69218-X

Bond, 1985, Ubiquitin is a heat shock protein in chicken embryo fibroblasts, Molec. cell. Biol., 5, 949

Bosch, 1991, The stress response in freshwater polyp Hydra, Heat Shock, 133, 10.1007/978-3-642-76679-4_15

Bosch, 1988, Thermotolerance and synthesis of heat shock proteins: These responses are present in Hydra attenuata but absent in Hydra oligactis, Proc. natn. Acad. Sci. U.S.A., 85, 7927, 10.1073/pnas.85.21.7927

Carlson, 1987, Microinjection of ubiquitin: changes in protein degradation in HeLa cells subjected to heat-shock, J. Cell Biol., 104, 547, 10.1083/jcb.104.3.547

Coleman, 1973, The oxygen consumption of Mytilus edulis in air, Comp. Biochem. Physiol., 45A, 393, 10.1016/0300-9629(73)90445-3

Craig, 1993, Chaperones: Helpers along the pathways to protein folding, Science, 260, 1902, 10.1126/science.8100364

Dayton, 1971, Competition, disturbance and community organization: The provision and subsequent utilization of space in a rocky intertidal community, Ecol. Monogr., 41, 351, 10.2307/1948498

De Zwaan, 1983, Carbohydrate catabolism in bivalves. In The Mollusca, Metabolic Biochemistry and Molecular Biomechanics, 137, 10.1016/B978-0-12-751401-7.50011-9

Dietz, 1992, The threshold induction temperature of the 90-kDa heat shock protein is subject to acclimatization in eurythermal goby fishes (genus Gillichthys), Proc. natn. Acad. Sci. U.S.A., 89, 3389, 10.1073/pnas.89.8.3389

Dill, 1990, Dominant forces in protein folding, Biochemistry, N.Y., 29, 7133, 10.1021/bi00483a001

Ellis, 1991, Molecular chaperones, A. Rev. Biochem., 60, 321, 10.1146/annurev.bi.60.070191.001541

Finley, 1991, Ubiquitination, A. Rev. Cell Biol., 7, 26, 10.1146/annurev.cb.07.110191.000325

Frydman, 1994, Folding of nascent polypeptide chains in a high molecular mass assembly with molecular chaperones, Nature, 370, 111, 10.1038/370111a0

Gething, 1992, Protein folding in the cell, Nature, 355, 33, 10.1038/355033a0

Haas, 1985, The immunochemical detection and quantification of intracellular ubiquitin–protein conjugates, J. biol. Chem., 260, 12464, 10.1016/S0021-9258(17)38895-6

Hahn, 1990, Thermotolerance, thermoresistance and thermosensitization, Stress Proteins In Biology and Medicine, 79

Hawkins, 1985, Relationships between the synthesis and breakdown of protein, dietary absorption and turnovers of nitrogen and carbon in the blue musssel, Mytilus edulis L, Oecologia, 66, 42, 10.1007/BF00378550

Hawkins, 1991, Protein turnover: a functional appraisal, Funct. Ecol., 5, 222, 10.2307/2389260

Hershko, 1992, The ubiquitin system for protein degradation, A. Rev. Biochem., 61, 761, 10.1146/annurev.bi.61.070192.003553

Hoffmann, 1991, Evolutionary Genetics and Environmental Stress. Oxford: Oxford University Press

Houlihan, 1991, Protein turnover in ectotherms and its relationships to energetics, Adv. comp. env. Physiol., 7, 1, 10.1007/978-3-642-75897-3_1

Huey, 1991, Physiological consequences of habitat selection, Am. Nat., 137, S91, 10.1086/285141

Huey, 1990, Physiological adjustments to fluctuating thermal environments: an ecological and evolutionary perspective, Stress Proteins in Biology and Medecine, 37

Huey, 1993, Evolution of resistance to high temperatures in ectotherms, Am. Nat., 142, S21, 10.1086/285521

Jaenicke, 1991, Protein stability and molecular adaptation to extreme conditions, Eur. J. Biochem., 202, 715, 10.1111/j.1432-1033.1991.tb16426.x

Kurtz, 1986, An ancient developmental induction: Heat-shock proteins induced in sporulation and oogenesis, Science, 231, 1154, 10.1126/science.3511530

Laemmli, 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227, 680, 10.1038/227680a0

Langer, 1992, Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding, Nature, 356, 683, 10.1038/356683a0

Lenski, 1993, Evolutionary response of Escherichia coli to thermal stress, Am. Nat., 142, S47, 10.1086/285522

Li, 1991, Thermal response of rat fibroblasts stably transfected with the human 70-kDa heat shock protein-encoding gene, Proc. natn. Acad. Sci. U.S.A., 88, 1681, 10.1073/pnas.88.5.1681

Lindquist, 1986, The heat-shock response, A. Rev. Biochem., 55, 1151, 10.1146/annurev.bi.55.070186.005443

Martin, 1991, Chaperonin-mediated protein folding at the surface of groEL through a “molten-globule”-like intermediate, Nature, 352, 36, 10.1038/352036a0

Mcdonald, 1988, The mussels Mytilus galloprovincialis and M. trossulus on the Pacific coast of North America, Mar. Biol., 99, 111, 10.1007/BF00644984

Newell, 1979, Biology of Intertidal Animals. Faversham, Kent, UK: Marine Ecological Surveys, Ltd

Parag, 1987, Effect of heat shock on protein degradation in mammalian cells: involvement of the ubiquitin system, EMBO J., 6, 55, 10.1002/j.1460-2075.1987.tb04718.x

Parsell, 1993, The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins, A. Rev. Genet., 27, 437, 10.1146/annurev.ge.27.120193.002253

Parsell, 1993, The role of heat-shock proteins in thermotolerance, Phil. Trans. R. Soc. Lond. B, 339, 279, 10.1098/rstb.1993.0026

Parsons, 1993, The importance and consequences of stress in living and fossil populations: from life-history variation to evolutionary change, Am. Nat., 142, S5, 10.1086/285520

Pickart, 1991, Dynamics of ubiquitin pools in developing sea urchin embryos, Dev. Growth Differ., 33, 587, 10.1111/j.1440-169X.1991.00587.x

Rechsteiner, 1987, Ubiquitin-mediated pathways for intracellular proteolysis, A. Rev. Cell Biol., 3, 1, 10.1146/annurev.cb.03.110187.000245

Sanchez, 1990, HSP104 required for induced thermotolerance, Science, 248, 1112, 10.1126/science.2188365

Sanchez, 1992, Hsp104 is required for tolerance to many forms of stress, EMBO J., 11, 2357, 10.1002/j.1460-2075.1992.tb05295.x

Sanders, 1992, Persistence of the heat-shock response over time in a common Mytilus mussel, Molec. mar. Biol. Biotech., 1, 147

Schröder, 1993, DnaK, DnaJ and GrpE form a cellular chaperone machinery capable of repairing heat-induced protein, EMBO J., 12, 4137, 10.1002/j.1460-2075.1993.tb06097.x

Skowyra, 1990, The E. coli dnaK gene product, the hsp70 homolog, can reactivate heatinactivated RNA polymerase in an ATP hydrolysis-dependent manner, Cell, 62, 939, 10.1016/0092-8674(90)90268-J

Smith, 1985, Measurement of protein using bicinchoninic acid, Analyt. Biochem., 150, 76, 10.1016/0003-2697(85)90442-7

Solomon, 1991, Changes in Hsp70 alter thermotolerance and heat-shock regulation in Drosophila, New Biol., 3, 1106

Somero, 1995, Proteins and temperature, A. Rev. Physiol., 57, 43, 10.1146/annurev.ph.57.030195.000355

Suchanek, 1978, The ecology of Mytilus edulis L. in exposed rocky intertidal communities, J. exp. mar. Biol. Ecol., 31, 105, 10.1016/0022-0981(78)90139-9

Widdows, 1979, Physiological and biochemical responses of bivalve molluscs to exposure to air, Comp. Biochem. Physiol., 62A, 301, 10.1016/0300-9629(79)90060-4

Widdows, 1985, Physiological responses of Mytilus edulis and Cardium edule to aerial exposure, Mar. Biol., 85, 217, 10.1007/BF00393242

Thông tin
Thông tin xuất bản

Journal of Experimental Biology

Tập 198 Số 7

1509-1518

Thông tin tác giả