SHARPIN-mediated regulation of protein arginine methyltransferase 5 controls melanoma growth
Hironari Tamiya, … , Kazuhiro Iwai, Ze’ev A. Ronai
Hironari Tamiya, … , Kazuhiro Iwai, Ze’ev A. Ronai
Published December 11, 2017
Citation Information: J Clin Invest. 2018;128(1):517-530. https://doi.org/10.1172/JCI95410.
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Research Article Cell biology Oncology

SHARPIN-mediated regulation of protein arginine methyltransferase 5 controls melanoma growth

Abstract

SHARPIN, an adaptor for the linear ubiquitin chain assembly complex (LUBAC), plays important roles in NF-κB signaling and inflammation. Here, we have demonstrated a LUBAC-independent role for SHARPIN in regulating melanoma growth. We observed that SHARPIN interacted with PRMT5, a type II protein arginine methyltransferase, and increased its multiprotein complex and methyltransferase activity. Activated PRMT5 controlled the expression of the transcription factors SOX10 and MITF by SHARPIN-dependent arginine dimethylation and inhibition of the transcriptional corepressor SKI. Activation of PRMT5 by SHARPIN counteracted PRMT5 inhibition by methylthioadenosine, a substrate of methylthioadenosine phosphorylase, which is codeleted with cyclin-dependent kinase inhibitor 2A (CDKN2A) in approximately 15% of human cancers. Collectively, we identified a LUBAC-independent role for SHARPIN in enhancing PRMT5 activity that contributes to melanomagenesis through the SKI/SOX10 regulatory axis.

Authors

Hironari Tamiya, Hyungsoo Kim, Oleksiy Klymenko, Heejung Kim, Yongmei Feng, Tongwu Zhang, Jee Yun Han, Ayako Murao, Scott J. Snipas, Lucia Jilaveanu, Kevin Brown, Harriet Kluger, Hao Zhang, Kazuhiro Iwai, Ze’ev A. Ronai

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Figure 2

SHARPIN plays a role in melanoma growth.

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SHARPIN plays a role in melanoma growth. (A) Colony-forming efficiency (...
(A) Colony-forming efficiency (CFE) assay of melanoma cell lines expressing scrambled (Scr) or HOIP-, HOIL-1L–, or SHARPIN-specific shRNA. Cells were seeded at 103/well and incubated for 14 days. Upper panels show representative images on day 14, and lower graph shows quantification of CFE (ImageJ). KD efficiency of each gene was confirmed with immunoblotting (see Supplemental Figure 1D). (B) CFE assay of melanoma cell lines expressing scrambled or SHARPIN-specific shRNAs (nos. 1, 2, and 3). Cells were seeded at 2.5 × 103/well and analyzed as in D. (C) Growth of WM115 cells (4 × 106 cells) expressing scrambled or SHARPIN-specific shRNA after subcutaneous injection into female nude mice. Tumor volumes were measured at the indicated time points. Data are presented as mean ± SD. n = 8 mice/group. Statistical significance was calculated using 2-way ANOVA. (D) CFE assay of WM793 and WM35 cells treated as indicated in C except that SHARPIN was reexpressed (+ rescue) as indicated. Upper panel shows representative images of colonies on day 14 after seeding at 103/well. Lower graph shows CFE quantification on day 14. All quantitation data are presented as mean ± SD (n = 3). (A, B, D) Statistical significance was calculated using 1-way ANOVA and Dunnett’s test. *P < 0.05; **P < 0.005; ***P < 0.0005 (2-tailed Student’s t test). (A and B) Data are representative of 3 experiments.