Cell Stress and Chaperones

, Volume 24, Issue 6, pp 1137–1149 | Cite as

Identification of an Apis cerana cerana MAP kinase phosphatase 3 gene (AccMKP3) in response to environmental stress

  • Yuzhen Chao
  • Chen Wang
  • Haihong Jia
  • Na Zhai
  • Hongfang Wang
  • Baohua Xu
  • Han LiEmail author
  • Xingqi GuoEmail author
Original Paper


MAP kinase phosphatase 3 (MKP3), a member of the dual-specificity protein phosphatase (DUSP) superfamily, has been widely studied for its role in development, cancer, and environmental stress in many organisms. However, the functions of MKP3 in various insects have not been well studied, including honeybees. In this study, we isolated an MKP3 gene from Apis cerana cerana and explored the role of this gene in the resistance to oxidation. We found that AccMKP3 is highly conserved in different species and shares the closest evolutionary relationship with AmMKP3. We determined the expression patterns of AccMKP3 under various stresses. qRT-PCR results showed that AccMKP3 was highly expressed during the pupal stages and in adult muscles. We further found that AccMKP3 was induced in all the stress treatments. Moreover, we discovered that the enzymatic activities of peroxidase, superoxide dismutase, and catalase increased and that the expression levels of several antioxidant genes were affected after AccMKP3 was knocked down. Collectively, these results suggest that AccMKP3 may be associated with antioxidant processes involved in response to various environmental stresses.


MAP kinase phosphatase 3 Oxidative stress RNA interference Environmental stresses Apis cerana cerana 


Funding information

This work was financially supported by Funds of the National Natural Science Foundation of China (No. 31572470) and the Earmarked Fund for the China Agriculture Research System (No. CARS-44).

Supplementary material

12192_2019_1036_Fig8_ESM.png (99 kb)
Supplementary Fig. 1

(PNG 98.8 kb)

12192_2019_1036_MOESM1_ESM.tif (574 kb)
High Resolution Image (TIF 574 kb)


  1. Alaux C, Ducloz F, Crauser D, Le Conte Y (2010) Diet effects on honeybee immunocompetence. Biol Lett 6:562–565. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Ali A, Rashid MA, Huang QY, Lei CL (2017) Influence of UV-A radiation on oxidative stress and antioxidant enzymes in Mythimna separata (Lepidoptera: Noctuidae). Environ Sci Pollut Res Int 24:8392–8398. CrossRefPubMedGoogle Scholar
  3. Asghari MH, Moloudizargari M, Bahadar H, Abdollahi M (2017) A review of the protective effect of melatonin in pesticide-induced toxicity. Expert Opin Drug Metab Toxicol 13:545–554. CrossRefPubMedGoogle Scholar
  4. Bafana A, Khan F, Suguna K (2017) Structural and functional characterization of mercuric reductase from Lysinibacillus sphaericus strain G1. Biometals 30:809–819. CrossRefPubMedGoogle Scholar
  5. Burmeister C, Kai L, Heinick A, Hussein A, Domagalski M, Walter RD, Liebau E (2008) Oxidative stress in Caenorhabditis elegans: protective effects of the Omega class glutathione transferase (GSTO-1). FASEB J 22:343–354. CrossRefPubMedGoogle Scholar
  6. Camps M, Nichols A, Gillieron C, Antonsson B, Muda M, Chabert C et al (1998) Catalytic activation of the phosphatase MKP-3 by ERK2 mitogen-activated protein kinase. Science 280:1262–1265CrossRefGoogle Scholar
  7. Camps M, Nichols A, Arkinstall S (2000) Dual specificity phosphatases: a gene family for control of MAP kinase function. FASEB J 14:6–16CrossRefGoogle Scholar
  8. Chen X, Yao P, Chu X, Hao L, Guo X, Xu B (2015) Isolation of arginine kinase from Apis cerana cerana and its possible involvement in response to adverse stress. Cell Stress Chaperones 20:169–183. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Corona M, Robinson GE (2006) Genes of the antioxidant system of the honey bee: annotation and phylogeny. Insect Mol Biol 15:687–701. CrossRefPubMedPubMedCentralGoogle Scholar
  10. de Carvalho LP, de Melo EJT (2018) Further aspects of Toxoplasma gondii elimination in the presence of metals. Parasitol Res 117:1245–1256. CrossRefPubMedGoogle Scholar
  11. Emanuele S, D’Anneo A, Calvaruso G, Cernigliaro C, Giuliano M, Lauricella M (2018) The double-edged sword profile of redox signaling: oxidative events as molecular switches in the balance between cell physiology and cancer. Chem Res Toxicol 31:201–210. CrossRefPubMedGoogle Scholar
  12. Ericsson A, Kotarsky K, Svensson M, Sigvardsson M, Agace W (2006) Functional characterization of the CCL25 promoter in small intestinal epithelial cells suggests a regulatory role for caudal-related homeobox (Cdx) transcription factors. J Immunol 176:3642–3651CrossRefGoogle Scholar
  13. Farooq A, Chaturvedi G, Mujtaba S, Plotnikova O, Zeng L, Dhalluin C, Ashton R, Zhou MM (2001) Solution structure of ERK2 binding domain of MAPK phosphatase MKP-3: structural insights into MKP-3 activation by ERK2. Mol Cell 7:387–399CrossRefGoogle Scholar
  14. Furukawa T, Horii A (2004) Molecular pathology of pancreatic cancer: in quest of tumor suppressor genes. Pancreas 28:253–256CrossRefGoogle Scholar
  15. Geng XM, Liu X, Ji M, Hoffmann WA, Grunden A, Xiang QY (2016) Enhancing heat tolerance of the little dogwood cornus canadensis L. f. with introduction of a superoxide reductase gene from the hyperthermophilic archaeon Pyrococcus furiosus. Front. Plant Sci 7:26. CrossRefGoogle Scholar
  16. Graves JA, Metukuri M, Scott D, Rothermund K, Prochownik EV (2009) Regulation of reactive oxygen species homeostasis by peroxiredoxins and c-Myc. J Biol Chem 284:6520–6529. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Gray EM (2013) Thermal acclimation in a complex life cycle: the effects of larval and adult thermal conditions on metabolic rate and heat resistance in Culex pipiens (Diptera: Culicidae). J Insect Physiol 59:1001–1007. CrossRefPubMedGoogle Scholar
  18. Harrison JF, Fewell JH (2002) Environmental and genetic influences on flight metabolic rate in the honey bee, Apis mellifera. Comp Biochem Physiol A Mol Integr Physiol 133:323–333CrossRefGoogle Scholar
  19. Hayes JD, Mcmahon M (2009) NRF2 and KEAP1 mutations: permanent activation of an adaptive response in cancer. Trends Biochem Sci 34:176–188. CrossRefPubMedGoogle Scholar
  20. Jeong DG et al (2014) The family-wide structure and function of human dual-specificity protein phosphatases. Acta Crystallogr D Biol Crystallogr 70:421–435. CrossRefPubMedGoogle Scholar
  21. Jia H, Ma M, Zhai N, Liu Z, Wang H, Guo X, Xu B (2017) Roles of a mitochondrial AccSCO2 gene from Apis cerana cerana in oxidative stress responses. J Inorg Biochem 175:9–19. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Karkali K, Panayotou G (2012) The Drosophila DUSP puckered is phosphorylated by JNK and p38 in response to arsenite-induced oxidative stress. Biochem Biophys Res Commun 418:301–306. CrossRefPubMedGoogle Scholar
  23. Kim SH et al (2002) Isolation and characterization of a Drosophila homologue of mitogen-activated protein kinase phosphatase-3 which has a high substrate specificity towards extracellular-signal-regulated kinase. Biochem J 361:143–151CrossRefGoogle Scholar
  24. Koga S, Kojima S, Kishimoto T, Kuwabara S, Yamaguchi A (2012) Over-expression of map kinase phosphatase-1 (MKP-1) suppresses neuronal death through regulating JNK signaling in hypoxia/re-oxygenation. Brain Res 1436:137–146. CrossRefPubMedGoogle Scholar
  25. Lee SB, Shin JS, Han HS, Lee HH, Park JC, Lee KT (2018) Kaempferol 7-O-beta-D-glucoside isolated from the leaves of Cudrania tricuspidata inhibits LPS-induced expression of pro-inflammatory mediators through inactivation of NF-κB, AP-1, and JAK-STAT in RAW 264.7 macrophages. Chem Biol Interact 284:101–111. CrossRefPubMedGoogle Scholar
  26. Li G, Zhao H, Wang H, Guo X, Guo X, Sun Q, Xu B (2016) Characterization of a decapentapletic gene (AccDpp) from Apis cerana cerana and its possible involvement in development and response to oxidative stress. PLoS One 11:e0149117. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Li C et al (2018) BHDPC is a novel neuroprotectant that provides anti-neuroinflammatory and neuroprotective effects by inactivating NF-κB and activating PKA/CREB. Front Pharmacol 9:614. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Liu S, Sun JP, Zhou B, Zhang ZY (2006) Structural basis of docking interactions between ERK2 and MAP kinase phosphatase 3. Proc Natl Acad Sci U S A 103:5326–5331. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408. CrossRefGoogle Scholar
  30. Lushchak VI (2011) Environmentally induced oxidative stress in aquatic animals. Aquat Toxicol 101:13–30. CrossRefPubMedGoogle Scholar
  31. Mates JM, Segura JA, Alonso FJ, Marquez J (2012) Oxidative stress in apoptosis and cancer: an update. Arch Toxicol 86:1649–1665. CrossRefPubMedGoogle Scholar
  32. Matsumura T, Matsumoto H, Hayakawa Y (2017) Heat stress hardening of oriental armyworms is induced by a transient elevation of reactive oxygen species during sublethal stress. Arch Insect Biochem Physiol:96. CrossRefGoogle Scholar
  33. Mazaira GI, Daneri-Becerra C, Zgajnar NR, Lotufo CM, Galigniana MD (2018) Gene expression regulation by heat-shock proteins: the cardinal roles of HSF1 and Hsp90. Biochem Soc Trans 46:51–65. CrossRefPubMedGoogle Scholar
  34. Molnar C, de Celis JF (2013) Tay bridge is a negative regulator of EGFR signalling and interacts with Erk and Mkp3 in the Drosophila melanogaster wing. PLoS Genet 9:e1003982. CrossRefPubMedPubMedCentralGoogle Scholar
  35. Moskovitz J, Oien DB (2010) Protein carbonyl and the methionine sulfoxide reductase system. Antioxid Redox Signal 12:405–415. CrossRefPubMedGoogle Scholar
  36. Muda M et al (1996) The dual specificity phosphatases M3/6 and MKP-3 are highly selective for inactivation of distinct mitogen-activated protein kinases. J Biol Chem 271:27205–27208CrossRefGoogle Scholar
  37. Muda M et al (1998) The mitogen-activated protein kinase phosphatase-3 N-terminal noncatalytic region is responsible for tight substrate binding and enzymatic specificity. J Biol Chem 273:9323–9329. CrossRefPubMedGoogle Scholar
  38. Oehrl W, Cotsiki M, Panayotou G (2013) Differential regulation of M3/6 (DUSP8) signaling complexes in response to arsenite-induced oxidative stress. Cell Signal 25:429–438. CrossRefPubMedGoogle Scholar
  39. Palacios C, Collins MK, Perkins GR (2001) The JNK phosphatase M3/6 is inhibited by protein-damaging stress. Curr Biol 11:1439–1443CrossRefGoogle Scholar
  40. Patterson KI, Brummer T, O’Brien PM, Daly RJ (2009) Dual-specificity phosphatases: critical regulators with diverse cellular targets. Biochem J 418:475–489CrossRefGoogle Scholar
  41. Ratnieks FLW (2006) Asian honey bees: biology, conservation and human interactions. Nature 442:249–249CrossRefGoogle Scholar
  42. Schatzman SS, Culotta VC (2018) Chemical warfare at the microorganismal level: a closer look at the superoxide dismutase enzymes of pathogens. ACS Infect Dis 4:893–903. CrossRefPubMedPubMedCentralGoogle Scholar
  43. Sharda PR, Bonham CA, Mucaki EJ, Butt Z, Vacratsis PO (2009) The dual-specificity phosphatase hYVH1 interacts with Hsp70 and prevents heat-shock-induced cell death. Biochem J 418:391–1401. CrossRefPubMedGoogle Scholar
  44. Shi W, Sun J, Xu B, Li H (2013) Molecular characterization and oxidative stress response of a cytochrome P450 gene (CYP4G11) from Apis cerana cerana. Z Naturforsch C 68:509–521CrossRefGoogle Scholar
  45. Stewart AE, Dowd S, Keyse SM, McDonald NQ (1999) Crystal structure of the MAPK phosphatase Pyst1 catalytic domain and implications for regulated activation. Nat Struct Biol 6:174–181. CrossRefPubMedGoogle Scholar
  46. Takagaki K, Shima H, Tanuma N, Nomura M, Satoh T, Watanabe M, Kikuchi K (2007) Characterization of a novel low-molecular-mass dual specificity phosphatase-4 (LDP-4) expressed in brain. Mol Cell Biochem 296:177–184. CrossRefPubMedGoogle Scholar
  47. Tang T, Huang DW, Zhou CQ, Li X, Xie QJ, Liu FS (2012) Molecular cloning and expression patterns of copper/zinc superoxide dismutase and manganese superoxide dismutase in Musca domestica. Gene 505:211–220. CrossRefPubMedPubMedCentralGoogle Scholar
  48. Wong VC et al (2012) Tumor suppressor dual-specificity phosphatase 6 (DUSP6) impairs cell invasion and epithelial-mesenchymal transition (EMT)-associated phenotype. Int J Cancer 130:83–95. CrossRefPubMedGoogle Scholar
  49. Zhang YY, Wu JW, Wang ZX (2011) Mitogen-activated protein kinase (MAPK) phosphatase 3-mediated cross-talk between MAPKs ERK2 and p38alpha. J Biol Chem 286:16150–16162. CrossRefPubMedPubMedCentralGoogle Scholar
  50. Zhao G, Wang C, Wang H, Gao L, Liu Z, Xu B, Guo X (2018) Characterization of the CDK5 gene in Apis cerana cerana (AccCDK5) and a preliminary identification of its activator gene, AccCDK5r1. Cell Stress Chaperones 23:13–28. CrossRefPubMedGoogle Scholar
  51. Zhu M, Zhang W, Liu F, Chen X, Li H, Xu B (2016) Characterization of an Apis cerana cerana cytochrome P450 gene (AccCYP336A1) and its roles in oxidative stresses responses. Gene 584:120–128. CrossRefPubMedGoogle Scholar

Copyright information

© Cell Stress Society International 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Crop Biology, College of Life SciencesShandong Agricultural UniversityTai’anPeople’s Republic of China
  2. 2.College of Animal Science and TechnologyShandong Agricultural UniversityTai’anPeople’s Republic of China

Personalised recommendations