Abstract
African clawed frogs (Xenopus laevis) endure bouts of severe drought in their natural habitats and survive the loss of approximately 30% of total body water due to dehydration. To investigate molecular mechanisms employed by X. laevis during periods of dehydration, the heat shock protein response, a vital component of the cytoprotective stress response, was characterized. Using western immunoblotting and multiplex technology, the protein levels of HSP27, HSP40, HSP60, HSP70, HSC70, and HSP90 were quantified in the liver, skeletal muscle, kidney, lung, and testes from control frogs and those that underwent medium or high dehydration (~16 or ~30% loss of total body water). Dehydration increased HSP27 (1.45–1.65-fold) in the kidneys and lungs, and HSP40 (1.39–2.50-fold) in the liver, testes, and skeletal muscle. HSP60 decreased in response to dehydration (0.43–0.64 of control) in the kidneys and lungs. HSP70 increased in the liver, lungs, and testes (1.39–1.70-fold) during dehydration, but had a dynamic response in the kidneys (levels increased 1.57-fold with medium dehydration, but decreased to 0.56 of control during high dehydration). HSC70 increased in the liver and kidneys (1.20–1.36-fold), but decreased in skeletal muscle (0.27–0.55 of control) during dehydration. Lastly, HSP90 was reduced in the kidney, lung, and skeletal muscle (0.39–0.69 of control) in response to dehydration, but rose in the testes (1.30-fold). Overall, the results suggest a dynamic tissue-specific heat shock protein response to whole body dehydration in X. laevis.
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References
Ali A, Salter-Cid L, Flajnik MF, Heikkila JJ (1996) Isolation and characterization of a cDNA encoding a Xenopus 70-kDa heat shock cognate protein, Hsc70.I. Comp Biochem Physiol B Biochem Mol Biol 113:681–687
Balinsky JB, Choritz EL, Coe CG, van der Schans GS (1967) Amino acid metabolism and urea synthesis in naturally aestivating Xenopus laevis. Comp Biochem Physiol 22:59–68
Brown D (2003) The ins and outs of aquaporin-2 trafficking. Am J Physiol Ren Physiol 284:F893–F901
Cajo GC, Horne BE, Kelley WL et al (2006) The role of the DIF motif of the DnaJ (Hsp40) co-chaperone in the regulation of the DnaK (Hsp70) chaperone cycle. J Biol Chem 281:12436–12444. doi:10.1074/jbc.M511192200
Cappello F, Conway de Macario E, Marasà L et al (2008) Hsp60 expression, new locations, functions and perspectives for cancer diagnosis and therapy. Cancer Biol Ther 7:801–809
Concannon CG, Orrenius S, Samali A (2001) Hsp27 inhibits cytochrome c-mediated caspase activation by sequestering both pro-caspase-3 and cytochrome c. Gene Expr 9:195–201
Garrido C, Bruey J-M, Ducasse C et al (2000) Hsp27 negatively regulates cell death by interacting with cytochrome c. Nat Cell Biol 2:645–652. doi:10.1038/35023595
Ghosh JC, Dohi T, Kang BH, Altieri DC (2008) Hsp60 regulation of tumor cell apoptosis. J Biol Chem 283:5188–5194. doi:10.1074/jbc.M705904200
Goldfarb SB, Kashlan OB, Watkins JN et al (2006) Differential effects of Hsc70 and Hsp70 on the intracellular trafficking and functional expression of epithelial sodium channels. Proc Natl Acad Sci U S A 103:5817–5822. doi:10.1073/pnas.0507903103
Greene MK, Maskos K, Landry SJ (1998) Role of the J-domain in the cooperation of Hsp40 with Hsp70. Proc Natl Acad Sci U S A 95:6108–6113
Heikkila JJ (2010) Heat shock protein gene expression and function in amphibian model systems. Comp Biochem Physiol A Mol Integr Physiol 156:19–33. doi:10.1016/j.cbpa.2010.01.024
Hillman SS (1978) The roles of oxygen delivery and electrolyte levels in the dehydrational death of Xenopus laevis. J Comp Physiol B 128:169–175. doi:10.1007/BF00689481
Jha KN, Coleman AR, Wong L et al (2013) Heat shock protein 90 functions to stabilize and activate the testis-specific serine/threonine kinases, a family of kinases essential for male fertility. J Biol Chem 288:16308–16320. doi:10.1074/jbc.M112.400978
Kampinga HH, Craig EA (2010) The HSP70 chaperone machinery: J proteins as drivers of functional specificity. Nat Rev Mol Cell Biol 11:579–592. doi:10.1038/nrm2941
Logan SM, Luu BE, Storey KB (2016) Turn down genes for WAT? Activation of anti-apoptosis pathways protects white adipose tissue in metabolically depressed thirteen-lined ground squirrels. Mol Cell Biochem 416:47–62. doi:10.1007/s11010-016-2695-0
Lu HAJ, Sun TX, Matsuzaki T et al (2007) Heat shock protein 70 interacts with aquaporin-2 and regulates its trafficking. J Biol Chem 282:28721–28732. doi:10.1074/jbc.M611101200
Luu BE, Storey KB (2015) Dehydration triggers differential microRNA expression in Xenopus laevis brain. Gene 573:64–69
Luu BE, Tessier SN, Duford DL, Storey KB (2015) The regulation of troponins I, C and ANP by GATA4 and Nkx2-5 in heart of hibernating thirteen-lined ground squirrels, Ictidomys tridecemlineatus. PLoS One 10:e0117747. doi:10.1371/journal.pone.0117747
Malik AI, Storey KB (2009a) Activation of extracellular signal-regulated kinases during dehydration in the African clawed frog, Xenopus laevis
Malik AI, Storey KB (2009b) Activation of antioxidant defense during dehydration stress in the African clawed frog. Gene 442:99–107. doi:10.1016/j.gene.2009.04.007
Malik AI, Storey KB (2011) Transcriptional regulation of antioxidant enzymes by FoxO1 under dehydration stress. Gene 485:114–119. doi:10.1016/j.gene.2011.06.014
Pearl EJ, Barker D, Day RC, Beck CW (2008) Identification of genes associated with regenerative success of Xenopus laevis hindlimbs. BMC Dev Biol 8:66. doi:10.1186/1471-213X-8-66
Samali A, Cai J, Zhivotovsky B et al (1999) Presence of a pre-apoptotic complex of pro-caspase-3, Hsp60 and Hsp10 in the mitochondrial fraction of Jurkat cells. EMBO J 18:2040–2048. doi:10.1093/emboj/18.8.2040
Storey KB, Storey JM (2011) Heat shock proteins and hypometabolism: adaptive strategy for proteome preservation. Res Rep Biol 2:57–68. doi:10.2147/RRB.S13351
Storey KB, Storey JM (2012) Aestivation: signaling and hypometabolism. J Exp Biol 215:1425–1433
Suh WC, Burkholder WF, Lu CZ et al (1998) Interaction of the Hsp70 molecular chaperone, DnaK, with its cochaperone DnaJ. Proc Natl Acad Sci U S A 95:15223–15228
Tuttle AM, Gauley J, Chan N, Heikkila JJ (2007) Analysis of the expression and function of the small heat shock protein gene, hsp27, in Xenopus laevis embryos. Comp Biochem Physiol A Mol Integr Physiol 147:112–121. doi:10.1016/j.cbpa.2006.12.003
Wijenayake S, Storey KB (2016) The role of DNA methylation during anoxia tolerance in a freshwater turtle (Trachemys scripta elegans). J Comp Physiol B 186:333–342. doi:10.1007/s00360-016-0960-x
Wolffe AP, Glover JF, Tata JR (1984) Culture shock. Synthesis of heat-shock-like proteins in fresh primary cell cultures. Exp Cell Res 154:581–590. doi:10.1016/0014-4827(84)90182-4
Wu CW, Biggar KK, Storey KB (2013) Dehydration mediated microRNA response in the African clawed frog Xenopus laevis. Gene 529:269–275. doi:10.1016/j.gene.2013.07.064
Xanthoudakis S, Roy S, Rasper D et al (1999) Hsp60 accelerates the maturation of pro-caspase-3 by upstream activator proteases during apoptosis. EMBO J 18:2049–2056. doi:10.1093/emboj/18.8.2049
Xu B, Hao Z, Jha KN et al (2008) Targeted deletion of Tssk1 and 2 causes male infertility due to haploinsufficiency. Dev Biol 319:211–222. doi:10.1016/j.ydbio.2008.03.047
Zou J, Guo Y, Guettouche T et al (1998) Repression of heat shock transcription factor HSF1 activation by HSP90 (HSP90 complex) that forms a stress-sensitive complex with HSF1. Cell 94:471–480. doi:10.1016/S0092-8674(00)81588-3
Acknowledgements
We thank Jan Storey for the editorial review of this article. This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery grant (#6793) to KBS. BEL holds a NSERC Canada Graduate Research Scholarship, SW holds a postgraduate Queen Elizabeth II Graduate Scholarship in Science and Technology, and KBS holds the Canada Research Chair in Molecular Physiology.
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BEL and SW are co-first authors. BEL conceived, designed, performed the experiments, analyzed the data, and wrote the paper. SW conceived, designed, performed the experiments, analyzed the data, and wrote the paper. AIM conceived and performed some of the experiments. KBS contributed reagents, materials, and wrote the paper.
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Luu, B.E., Wijenayake, S., Malik, A.I. et al. The regulation of heat shock proteins in response to dehydration in Xenopus laevis . Cell Stress and Chaperones 23, 45–53 (2018). https://doi.org/10.1007/s12192-017-0822-9
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DOI: https://doi.org/10.1007/s12192-017-0822-9