Skip to main content

Advertisement

SpringerLink
Log in

Glucosamine abrogates the stem cell factor + endothelin-1-induced stimulation of melanogenesis via a deficiency in MITF expression due to the proteolytic degradation of CREB in human melanocytes

  • Original Paper
  • Published:
Archives of Dermatological Research Aims and scope Submit manuscript

Abstract

We have already reported that glucosamine (GlcN) distinctly abrogates the pigmentation of human epidermal equivalents stimulated by stem cell factor + endothelin-1 (SE). In this study, we characterized the molecular mechanism involved in the anti-melanogenic effects of GlcN using normal human melanocytes (NHMs) in culture. The SE-stimulated gene (12 h) and protein (24 h) expression levels of melanocyte-specific proteins (at the indicated times post-stimulation) were significantly abrogated by pretreatment with GlcN for 72 h. Western blotting analysis of the phosphorylation of intracellular signaling molecules in the MAPK pathway revealed that despite the significantly decreased level of total CREB protein at all times post-stimulation, the SE-stimulated phosphorylation of ERK, CREB and MITF is not attenuated at 15 min post-stimulation in GlcN-treated NHMs. However, the SE-stimulated protein expression level of total MITF at 2 and 6 h post-stimulation was significantly abrogated by 72 h pretreatment with GlcN. Consistently, pretreatment with GlcN for 72 h abrogated the stimulated gene and protein expression levels of MITF at 1 h and 2 h post-stimulation, respectively. Analysis of gene and protein expression levels also demonstrated that pretreatment with GlcN for 72 h significantly reduced the protein levels of CREB and MITF without affecting their gene expression levels prior to the SE stimulation. Silencing with a CREB siRNA distinctly abrogated the SE-stimulated expression of MITF (at 2 h post-stimulation) and melanocyte-specific proteins (at 24 h post-stimulation). Similarly, transfection of MITF siRNA markedly abrogated the SE-stimulated expression of MITF protein and melanocyte-specific proteins at 2 and 24 h post-stimulation, respectively. Finally, the decreased levels of CREB and MITF proteins induced by 72 h pretreatment with GlcN were abrogated by the co-addition of the proteosomal degradation inhibitor MG132. These findings suggest that the anti-melanogenic effect elicited by GlcN is mediated via the decreased expression of MITF which results from the attenuated transcriptional activity of CREB due to proteolytic degradation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

GlcN:

Glucosamine

EDN:

Endothelin

SCF:

Stem cell factor

MITF:

Microphthalmia-associated transcription factor

CREB:

Cyclic AMP responsive element binding protein

NHMs:

Normal human melanocytes

TYR:

Tyrosinase

TRP1:

Tyrosinase-related protein-1

TRP2:

Tyrosinase-related protein-2 (or dopachrome tautomerase)

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HEEs:

Human epidermal equivalents

References

  1. Koch HU, Schwarz RT, Scholtissek C (1979) Glucosamine itself mediates reversible inhibition of protein glycosylation. A study of glucosamine metabolism at inhibitory concentrations in influenza-virus-infected cells. Eur J Biochem 94(2):515–522

    Article  PubMed  CAS  Google Scholar 

  2. Nakamura K, Compans RW (1978) Effects of glucosamine, 2-deoxyglucose, and tunicamycin on glycosylation, sulfation, and assembly of influenza viral proteins. Virology 84(2):303–319

    Article  PubMed  CAS  Google Scholar 

  3. Suh HN, Lee YJ, Kim MO, Ryu JM, Han HJ (2014) Glucosamine-induced Sp1 O-GlcNAcylation ameliorates hypoxia-induced SGLT dysfunction in primary cultured renal proximal tubule cells. J Cell Physiol 229(10):1557–1568

    Article  PubMed  CAS  Google Scholar 

  4. Imokawa G, Mishima Y (1982) Loss of melanogenic properties in tyrosinases induced by glucosylation inhibitors within malignant melanoma cells. Cancer Res 42(5):1994–2002

    PubMed  CAS  Google Scholar 

  5. Imokawa G, Mishima Y (1984) Functional analysis of tyrosinase isozymes of cultured malignant melanoma cells during the recovery period following interrupted melanogenesis induced by glycosylation inhibitors. J Invest Dermatol 83(3):196–201

    Article  PubMed  CAS  Google Scholar 

  6. Imokawa G, Mishima Y (1985) Analysis of tyrosinases as asparagin-linked oligosaccharides by concanavalin A lectin chromatography: appearance of new segment of tyrosinases in melanoma cells following interrupted melanogenesis induced by glycosylation inhibitors. J Invest Dermatol 85(2):165–168

    Article  PubMed  CAS  Google Scholar 

  7. Imokawa G, Mishima Y (1986) Importance of glycoproteins in the initiation of melanogenesis: an electron microscopic study of B-16 melanoma cells after release from inhibition of glycosylation. J Invest Dermatol 87(3):319–325

    Article  PubMed  CAS  Google Scholar 

  8. Slominski A, Tobin DJ, Shibahara S, Wortsman J (2004) Melanin pigmentation in mammalian skin and its hormonal regulation. Physiol Rev 84(4):1155–1228

    Article  PubMed  CAS  Google Scholar 

  9. Imokawa G, Ishida K (2014) Inhibitors of intracellular signaling pathways that lead to stimulated epidermal pigmentation: perspective of anti-pigmenting agents. Int J Mol Sci 15(5):8293–8315

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Slominski A, Zmijewski MA, Pawelek J (2012) L-tyrosine and L-dihyroxyphenylalanine as hormone-like regulators of melanocyte functions. Pigment Cell Melanoma Res 25(1):14–27

    Article  PubMed  CAS  Google Scholar 

  11. Wakabayashi Y, Nakajima H, Imokawa G (2013) Abrogating effect of N-linked carbohydrate modifiers on the stem cell factor and endothelin-1-stimulated epidermal pigmentation in human epidermal equivalents. J Dermatol Sci 69(3):215–228

    Article  PubMed  CAS  Google Scholar 

  12. Bentley NJ, Eisen T, Goding CR (1994) Melanocyte-specific expression of the human tyrosinase promoter: activation by the microphthalmia gene product and role of the initiator. Mol Cell Biol 14(12):7996–8006

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Bertolotto C, Buscà R, Abbe P, Bille K, Aberdam E, Ortonne JP, Ballotti R (1998) Different cis-acting elements are involved in the regulation of TRP1 and TRP2 promoter activities by cyclic AMP: pivotal role of M boxes (GTCATGTGCT) and of microphthalmia. Mol Cell Biol 18(2):694–702

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  14. Du J, Miller AJ, Widlund HR, Horstmann MA, Ramaswamy S, Fisher DE (2003) MLANA/MART1 and SILV/PMEL17/GP100 are transcriptionally regulated by MITF in melanocytes and melanoma. Am J Pathol 163(1):333–343

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Sato-Jin K, Nishimura EK, Akasaka E, Huber W, Nakano H, Miller A, Du J, Wu M, Hanada K, Sawamura D, Fisher DE, Imokawa G (2008) Epistatic connections between microphthalmia-associated transcription factor and endothelin signaling in Waardenburg syndrome and other pigmentary disorders. FASEB J 22(4):1155–1168

    Article  PubMed  CAS  Google Scholar 

  16. Nakajima H, Wakabayashi Y, Wakamatsu K, Imokawa G (2011) An extract of Melia toosendan attenuates endothelin-1-stimulated pigmentation in human epidermal equivalents through the interruption of PKC activity within melanocytes. Arch Dermatol Res 303(4):263–276

    Article  PubMed  Google Scholar 

  17. Nakajima H, Fukazawa K, Wakabayashi Y, Wakamatsu K, Imokawa G (2012) Withania somnifera extract attenuates stem cell factor-stimulated pigmentation in human epidermal equivalents through interruption of ERK phosphorylation within melanocytes. J Nat Med 66(3):435–446

    Article  PubMed  Google Scholar 

  18. Nakajima H, Fukazawa K, Wakabayashi Y, Wakamatsu K, Senda K, Imokawa G (2012) Abrogating effect of a xanthophyll carotenoid astaxanthin on the stem cell factor-induced stimulation of human epidermal pigmentation. Arch Dermatol Res 304(10):803–816

    Article  PubMed  CAS  Google Scholar 

  19. Mizutani Y, Hayashi N, Kawashima M, Imokawa G (2010) A single UVB exposure increases the expression of functional KIT in human melanocytes by up-regulating MITF expression through the phosphorylation of p38/CREB. Arch Dermatol Res 302(4):283–294

    Article  PubMed  CAS  Google Scholar 

  20. Newton RA, Cook AL, Roberts DW, Leonard JH, Sturm RA (2007) Post-transcriptional regulation of melanin biosynthetic enzymes by cAMP and resveratrol in human melanocytes. J Invest Dermatol 127(9):2216–2227

    Article  PubMed  CAS  Google Scholar 

  21. Imokawa G, Kobayasi T, Miyagishi M (2000) Intracellular signaling mechanisms leading to synergistic effects of endothelin-1 and stem cell factor on proliferation of cultured human melanocytes. Cross-talk via trans-activation of the tyrosine kinase c-kit receptor. J Biol Chem 275(43):33321–33328

    Article  PubMed  CAS  Google Scholar 

  22. Potterf SB, Furumura M, Dunn KJ, Arnheiter H, Pavan WJ (2000) Transcription factor hierarchy in Waardenburg syndrome: regulation of MITF expression by SOX10 and PAX3. Hum Genet 107(1):1–6

    Article  PubMed  CAS  Google Scholar 

  23. Wan P, Hu Y, He L (2011) Regulation of melanocyte pivotal transcription factor MITF by some other transcription factors. Mol Cell Biochem 354(1–2):241–246

    Article  PubMed  CAS  Google Scholar 

  24. Mascarenhas JB, Littlejohn EL, Wolsky RJ, Young KP, Nelson M, Salgia R, Lang D (2010) PAX3 and SOX10 activate MET receptor expression in melanoma. Pigment Cell Melanoma Res 23(2):225–237

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Guan D, Wang H, Li VE, Xu Y, Yang M, Shen Z (2009) N-glycosylation of ATF6beta is essential for its proteolytic cleavage and transcriptional repressor function to ATF6alpha. J Cell Biochem 108(4):825–831

    Article  PubMed  CAS  Google Scholar 

  26. Azuma Y, Miura K, Higai K, Matsumoto K (2007) Protein O-N-acetylglucosaminylation modulates promoter activities of cyclic AMP response element and activator protein 1 and enhances E-selectin expression on HuH-7 human hepatoma cells. Biol Pharm Bull 30(12):2284–2289

    Article  PubMed  CAS  Google Scholar 

  27. Guinez C, Mir AM, Dehennaut V, Cacan R, Harduin-Lepers A, Michalski JC, Lefebvre T (2008) Protein ubiquitination is modulated by O-GlcNAc glycosylation. FASEB J 22(8):2901–2911

    Article  PubMed  CAS  Google Scholar 

  28. Han I, Kudlow JE (1997) Reduced O glycosylation of Sp1 is associated with increased proteasome susceptibility. Mol Cell Biol 17(5):2550–2558

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

Funding

There is no funding source.

Author information

Author notes

  1. Takao Niwano and Shuko Terazawa contributed equally to this work.

Authors and Affiliations

  1. Center for Bioscience Research and Education, Utsunomiya University, 350 Mine Utsunomiya, Tochigi, 321-8505, Japan

    Takao Niwano, Shuko Terazawa, Yoriko Sato & Genji Imokawa

  2. School of Bioscience and Technology, Tokyo University of Technology, Tokyo, Japan

    Takahiro Kato & Hiroaki Nakajima

  3. Research Institute for Biological Functions, Chubu University, Aichi, Japan

    Genji Imokawa

Authors
  1. Takao Niwano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuko Terazawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoriko Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takahiro Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hiroaki Nakajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Genji Imokawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Genji Imokawa.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 217 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Niwano, T., Terazawa, S., Sato, Y. et al. Glucosamine abrogates the stem cell factor + endothelin-1-induced stimulation of melanogenesis via a deficiency in MITF expression due to the proteolytic degradation of CREB in human melanocytes. Arch Dermatol Res 310, 625–637 (2018). https://doi.org/10.1007/s00403-018-1850-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00403-018-1850-8

Keywords

Search

Navigation