Unphosphorylated HSP27 (HSPB1) regulates the translation initiation process via a direct association with eIF4E in osteoblasts

Kuroyanagi,Gen; Tokuda,Haruhiko; Yamamoto,Naohiro; Matsushima‑Nishiwaki,Rie; Kozawa,Osamu; Otsuka,Takanobu

doi:10.3892/ijmm.2015.2274

Unphosphorylated HSP27 (HSPB1) regulates the translation initiation process via a direct association with eIF4E in osteoblasts

Authors:
- Gen Kuroyanagi
- Haruhiko Tokuda
- Naohiro Yamamoto
- Rie Matsushima‑Nishiwaki
- Osamu Kozawa
- Takanobu Otsuka
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Affiliations: Department of Orthopedic Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi 467‑8601, Japan, Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu 501‑1194, Japan
Published online on: July 7, 2015 https://doi.org/10.3892/ijmm.2015.2274
Pages: 881-889

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Abstract

Heat-shock protein 27 (HSP27/HSPB1) and its phosphorylation are implicated in multiple physiological and pathophysiological cell functions. Our previous study reported that unphosphorylated HSP27 has an inhibitory role in triiodothyronine (T3)‑induced osteocalcin (OC) synthesis in osteoblasts. However, the mechanisms behind the HSP27‑mediated effects on osteoblasts remain to be clarified. In the present study, to investigate the exact mechanism of HSP27 and its phosphorylation in osteoblasts, the molecular targets of HSP27 were explored using osteoblast‑like MC3T3‑E1 cells. The levels of OC mRNA induced by T3 in the HSP27‑overexpressing cells did not show any significant differences compared with those in the control empty vector‑transfected cells. Therefore, the interactions between HSP27 and translational molecules were focused on, including eukaryotic translation initiation factor 4E (eIF4E), eIF4G and 4E‑binding protein 1 (4E‑BP1). The HSP27 protein in the unstimulated cells co‑immunoprecipitated with eIF4E, but not eIF4G or 4E‑BP1. In addition, the association of eIF4E with 4E‑BP1 was observed in the HSP27‑overexpressing cells, as well as in the control cells. Under T3 stimulation, the binding of eIF4E to eIF4G was markedly attenuated in the HSP27‑overexpressing cells compared with the control cells. In addition, the binding of HSP27 to eIF4E in the unstimulated cells was diminished by the phosphorylation of HSP27. In response to T3 stimulation, the association of eIF4E with eIF4G in the unphosphorylatable HSP27‑overexpressing cells was markedly reduced compared with the phospho‑mimic HSP27‑overexpressing cells. Taken together, these findings strongly suggest that unphosphorylated HSP27 associates with eIF4E in osteoblasts and suppresses the translation initiation process.

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1	Mymrikov EV, Seit-Nebi AS and Gusev NB: Large potentials of small heat shock proteins. Physiol Rev. 91:1123–1159. 2011. View Article : Google Scholar : PubMed/NCBI
2	Kampinga HH, Hageman J, Vos MJ, Kubota H, Tanguay RM, Bruford EA, Cheetham ME, Chen B and Hightower LE: Guidelines for the nomenclature of the human heat shock proteins. Cell Stress Chaperones. 14:105–111. 2009. View Article : Google Scholar :
3	Kriehuber T, Rattei T, Weinmaier T, Bepperling A, Haslbeck M and Buchner J: Independent evolution of the core domain and its flanking sequences in small heat shock proteins. FASEB J. 24:3633–3642. 2010. View Article : Google Scholar : PubMed/NCBI
4	Dubińska-Magiera M, Jabłońska J, Saczko J, Kulbacka J, Jagla T and Daczewska M: Contribution of small heat shock proteins to muscle development and function. FEBS Lett. 588:517–530. 2014. View Article : Google Scholar
5	Kostenko S and Moens U: Heat shock protein 27 phosphorylation: Kinases, phosphatases, functions and pathology. Cell Mol Life Sci. 66:3289–3307. 2009. View Article : Google Scholar : PubMed/NCBI
6	Landry J, Lambert H, Zhou M, Lavoie JN, Hickey E, Weber LA and Anderson CW: Human HSP27 is phosphorylated at serines 78 and 82 by heat shock and mitogen-activated kinases that recognize the same amino acid motif as S6 kinase II. J Biol Chem. 267:794–803. 1992.PubMed/NCBI
7	Hayes D, Napoli V, Mazurkie A, Stafford WF and Graceffa P: Phosphorylation dependence of hsp27 multimeric size and molecular chaperone function. J Biol Chem. 284:18801–18807. 2009. View Article : Google Scholar : PubMed/NCBI
8	Matsushima-Nishiwaki R, Takai S, Adachi S, Minamitani C, Yasuda E, Noda T, Kato K, Toyoda H, Kaneoka Y, Yamaguchi A, et al: Phosphorylated heat shock protein 27 represses growth of hepatocellular carcinoma via inhibition of extracellular signal-regulated kinase. J Biol Chem. 283:18852–18860. 2008. View Article : Google Scholar : PubMed/NCBI
9	Kular J, Tickner J, Chim SM and Xu J: An overview of the regulation of bone remodelling at the cellular level. Clin Biochem. 45:863–873. 2012. View Article : Google Scholar : PubMed/NCBI
10	Chim SM, Tickner J, Chow ST, Kuek V, Guo B, Zhang G, Rosen V, Erber W and Xu J: Angiogenic factors in bone local environment. Cytokine Growth Factor Rev. 24:297–310. 2013. View Article : Google Scholar : PubMed/NCBI
11	Uozaki H, Horiuchi H, Ishida T, Iijima T, Imamura T and Machinami R: Overexpression of resistance-related proteins (metallothioneins, glutathione-S-transferase pi, heat shock protein 27, and lung resistance-related protein) in osteosarcoma. Relationship with poor prognosis Cancer. 79:2336–2344. 1997.
12	Tiffee JC, Griffin JP and Cooper LF: Immunolocalization of stress proteins and extracellular matrix proteins in the rat tibia. Tissue Cell. 32:141–147. 2000. View Article : Google Scholar : PubMed/NCBI
13	Leonardi R, Barbato E, Paganelli C and Lo Muzio L: Immunolocalization of heat shock protein 27 in developing jaw bones and tooth germs of human fetuses. Calcif Tissue Int. 75:509–516. 2004. View Article : Google Scholar
14	Shakoori AR, Oberdorf AM, Owen TA, Weber LA, Hickey E, Stein JL, Lian JB and Stein GS: Expression of heat shock genes during differentiation of mammalian osteoblasts and promyelocytic leukemia cells. J Cell Biochem. 48:277–287. 1992. View Article : Google Scholar : PubMed/NCBI
15	Kozawa O, Niwa M, Matsuno H, Ishisaki A, Kato K and Uematsu T: Stimulatory effect of basic fibroblast growth factor on induction of heat shock protein 27 in osteoblasts: Role of protein kinase C. Arch Biochem Biophys. 388:237–242. 2001. View Article : Google Scholar : PubMed/NCBI
16	Kozawa O, Niwa M, Matsuno H, Tokuda H, Miwa M, Ito H, Kato K and Uematsu T: Sphingosine 1-phosphate induces heat shock protein 27 via p38 mitogen-activated protein kinase activation in osteoblasts. J Bone Miner Res. 14:1761–1767. 1999. View Article : Google Scholar : PubMed/NCBI
17	Kato K, Adachi S, Matsushima-Nishiwaki R, Minamitani C, Natsume H, Katagiri Y, Hirose Y, Mizutani J, Tokuda H, Kozawa O, et al: Regulation by heat shock protein 27 of osteocalcin synthesis in osteoblasts. Endocrinology. 152:1872–1882. 2011. View Article : Google Scholar : PubMed/NCBI
18	Arrigo AP and Gibert B: HspB1, HspB5 and HspB4 in human cancers: Potent oncogenic role of some of their client proteins. Cancers (Basel). 6:333–365. 2014. View Article : Google Scholar
19	Gibert B, Eckel B, Fasquelle L, Moulin M, Bouhallier F, Gonin V, Mellier G, Simon S, Kretz-Remy C, Arrigo AP, et al: Knock down of heat shock protein 27 (HspB1) induces degradation of several putative client proteins. PLoS One. 7:e297192012. View Article : Google Scholar : PubMed/NCBI
20	Andrieu C, Taieb D, Baylot V, Ettinger S, Soubeyran P, De-Thonel A, Nelson C, Garrido C, So A, Fazli L, et al: Heat shock protein 27 confers resistance to androgen ablation and chemotherapy in prostate cancer cells through eIF4E. Oncogene. 29:1883–1896. 2010. View Article : Google Scholar : PubMed/NCBI
21	Kong J and Lasko P: Translational control in cellular and developmental processes. Nat Rev Genet. 13:383–394. 2012. View Article : Google Scholar : PubMed/NCBI
22	Gilbert RJ, Gordiyenko Y, von der Haar T, Sonnen AF, Hofmann G, Nardelli M, Stuart DI and McCarthy JE: Reconfiguration of yeast 40S ribosomal subunit domains by the translation initiation multifactor complex. Proc Natl Acad Sci USA. 104:5788–5793. 2007. View Article : Google Scholar : PubMed/NCBI
23	Sonenberg N and Gingras AC: The mRNA 5′ cap-binding protein eIF4E and control of cell growth. Curr Opin Cell Biol. 10:268–275. 1998. View Article : Google Scholar : PubMed/NCBI
24	Jia Y, Polunovsky V, Bitterman PB and Wagner CR: Cap-dependent translation initiation factor eIF4E: An emerging anticancer drug target. Med Res Rev. 32:786–814. 2012. View Article : Google Scholar : PubMed/NCBI
25	Kubisch C, Dimagno MJ, Tietz AB, Welsh MJ, Ernst SA, Brandt-Nedelev B, Diebold J, Wagner AC, Göke B, Williams JA, et al: Overexpression of heat shock protein Hsp27 protects against cerulein-induced pancreatitis. Gastroenterology. 127:275–286. 2004. View Article : Google Scholar : PubMed/NCBI
26	Sudo H, Kodama HA, Amagai Y, Yamamoto S and Kasai S: In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol. 96:191–198. 1983. View Article : Google Scholar : PubMed/NCBI
27	Kozawa O, Tokuda H, Miwa M, Kotoyori J and Oiso Y: Cross-talk regulation between cyclic AMP production and phosphoinositide hydrolysis induced by prostaglandin E₂ in osteoblast-like cells. Exp Cell Res. 198:130–134. 1992. View Article : Google Scholar : PubMed/NCBI
28	Matsushima-Nishiwaki R, Kumada T, Nagasawa T, Suzuki M, Yasuda E, Okuda S, Maeda A, Kaneoka Y, Toyoda H and Kozawa O: Direct association of heat shock protein 20 (HSPB6) with phosphoinositide 3-kinase (PI3K) in human hepatocellular carcinoma: Regulation of the PI3K activity. PLoS One. 8:e784402013. View Article : Google Scholar : PubMed/NCBI
29	Zhang W, Yang N and Shi XM: Regulation of mesenchymal stem cell osteogenic differentiation by glucocorticoid-induced leucine zipper (GILZ). J Biol Chem. 283:4723–4729. 2008. View Article : Google Scholar
30	Simpson DA, Feeney S, Boyle C and Stitt AW: Retinal VEGF mRNA measured by SYBR green I fluorescence: A versatile approach to quantitative PCR. Mol Vis. 6:178–183. 2000.PubMed/NCBI
31	Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI
32	Cuesta R, Laroia G and Schneider RJ: Chaperone hsp27 inhibits translation during heat shock by binding eIF4G and facilitating dissociation of cap-initiation complexes. Genes Dev. 14:1460–1470. 2000.PubMed/NCBI
33	Kato K, Tokuda H, Adachi S, Matsushima-Nishiwaki R, Yamauchi J, Natsume H, Minamitani C, Mizutani J, Otsuka T and Kozawa O: Role of heat shock protein 27 in transforming growth factor-β-stimulated vascular endothelial growth factor release in osteoblasts. Int J Mol Med. 27:423–428. 2011.PubMed/NCBI
34	Barutta F, Pinach S, Giunti S, Vittone F, Forbes JM, Chiarle R, Arnstein M, Perin PC, Camussi G, Cooper ME, et al: Heat shock protein expression in diabetic nephropathy. Am J Physiol Renal Physiol. 295:F1817–F1824. 2008. View Article : Google Scholar : PubMed/NCBI
35	Singh D, McCann KL and Imani F: MAPK and heat shock protein 27 activation are associated with respiratory syncytial virus induction of human bronchial epithelial monolayer disruption. Am J Physiol Lung Cell Mol Physiol. 293:L436–L445. 2007. View Article : Google Scholar : PubMed/NCBI