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Regulation of pro-opiomelanocortin (POMC) gene transcription by interleukin-31 via early growth response 1 (EGR-1) in HaCaT keratinocytes

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Abstract

Pro-opiomelanocortin (POMC) is a large precursor protein of and β-endorphin. POMC expressed in keratinocytes regulates various pathophysiological responses, such as pruritus in atopic dermatitis. Interleukin (IL)-31 is a T helper 2 (Th2)-derived cytokine that functions as a pruritogen, stimulating the sensory neurons in the skin. However, the regulatory mechanism underlying IL-31-induced POMC expression in keratinocytes remains largely unknown. Herein, using a 5′-serial deletion and site-specific mutation constructs of the regulatory region of POMC, we demonstrated that a putative EGR1-binding sequence (EBS) motif in POMC is required for its upregulation by IL-31 in HaCaT keratinocytes. Notably, EGR-1 directly interacted with the EBS motif in POMC. The ectopic expression of EGR-1 stimulated the POMC promoter activity, whereas the knockdown of EGR-1 expression by RNA interference reduced IL-31-induced POMC expression. Furthermore, we observed that three major mitogen-activated protein kinases, ERK, JNK, and p38 kinase, mediated IL-31-induced EGR-1 expression. In summary, our results suggest that EGR-1 trans-activates POMC in response to IL-31 stimulation in HaCaT keratinocytes.

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Abbreviations

ACTH:

Adrenocorticotropic hormone

ANOVA:

One-way analysis of variance

EGR1:

Early growth response 1

EBS:

EGR-1-binding sequence

EMSA:

Electrophoretic mobility shift assay

ERK1/2:

Extracellular signal-regulated kinase 1/2

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

IL-31:

Interleukin 31

MAPK:

Mitogen-activated protein kinase

MEK:

MAPK kinase

MSH:

Melanocyte-stimulating hormone

POMC:

Pro-opiomelanocortin

QR-PCR:

Quantitative real-time PCR

RT-PCR:

Reverse transcription-polymerase chain reaction

shRNA:

Short hairpin RNA

TSLP:

Thymic stromal lymphopoietin

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Acknowledgements

This study was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Science, ICT and Future Planning (Grant No. NRF-2018R1A2B2004653). This paper was supported by the KU Research Professor Program of Konkuk University.

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Authors and Affiliations

  1. Department of Biological Sciences, Sanghuh College of Life Sciences, Konkuk University, Seoul, 05029, Republic of Korea

    Hyunjin Yeo, Sung Shin Ahn, Young Han Lee & Soon Young Shin

  2. Cancer and Metabolism Institute, Konkuk University, Seoul, 05029, Republic of Korea

    Young Han Lee & Soon Young Shin

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  1. Hyunjin Yeo
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  2. Sung Shin Ahn
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HY constructed the promoter reports and performed the reporter assays. SSA performed the DNA-protein binding experiments and immunoblotting. YHL designed the experiments and analyzed the data. SYS conceived and supervised the study.

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Correspondence to Soon Young Shin.

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Yeo, H., Ahn, S.S., Lee, Y.H. et al. Regulation of pro-opiomelanocortin (POMC) gene transcription by interleukin-31 via early growth response 1 (EGR-1) in HaCaT keratinocytes. Mol Biol Rep 47, 5953–5962 (2020). https://doi.org/10.1007/s11033-020-05668-0

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