Uveal melanoma–associated mutations in PLCβ4 are constitutively activating and promote melanocyte proliferation and tumorigenesis

Phan, Hoa T.N.; Kim, Nam Hoon; Wei, Wenhui; Tall, Gregory G.; Smrcka, Alan V.

doi:10.1126/scisignal.abj4243

Cited by 11 publications

(8 citation statements)

References 62 publications

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“…The somatic substitution of glutamine 209 or arginine 183 in either GNAQ or GNA11 generates constitutive activity of the G protein and is a characteristic early event in uveal melanoma, occurring in up to 90% of cases ( Van Raamsdonk et al, 2010 ). A subset of cases without GNAQ or GNA11 mutations exhibits a gain-of-function hotspot mutation in PLCB4 ( Johansson et al, 2016 ; van de Nes et al, 2017 ; Phan et al, 2021 ). These mutations are also frequent in blue nevi and primary melanomas of the central nervous system ( Van Raamsdonk et al, 2010 ; Van Raamsdonk et al, 2009 ; Küsters-Vandevelde et al, 2010 ).…”

Section: Introductionmentioning

confidence: 99%

Crosstalk with keratinocytes causes GNAQ oncogene specificity in melanoma

Urtatiz

Haage

Tanentzapf

et al. 2021

eLife

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Different melanoma subtypes exhibit specific and non-overlapping sets of oncogene and tumor suppressor mutations, despite a common cell of origin in melanocytes. For example, activation of the Gαq/11 signaling pathway is a characteristic initiating event in primary melanomas that arise in the dermis, uveal tract or central nervous system. It is rare in melanomas arising in the epidermis. The mechanism for this specificity is unknown. Here, we present evidence that in the mouse, crosstalk with the epidermal microenvironment actively impairs the survival of melanocytes expressing the GNAQQ209L oncogene. We found that GNAQQ209L, in combination with signaling from the interfollicular epidermis (IFE), stimulates dendrite extension, leads to actin cytoskeleton disorganization, inhibits proliferation and promotes apoptosis in melanocytes. The effect was reversible and paracrine. In contrast, the epidermal environment increased the survival of wildtype and BrafV600E expressing melanocytes. Hence, our studies reveal the flip side of Gaq/11 signaling, which was hitherto unsuspected. In the future, the identification of the epidermal signals that restrain the GNAQQ209L oncogene could suggest novel therapies for GNAQ and GNA11 mutant melanomas.

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Section: Introductionmentioning

confidence: 99%

Crosstalk with keratinocytes causes GNAQ oncogene specificity in melanoma

Urtatiz

Haage

Tanentzapf

et al. 2021

eLife

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“…Gel electrophoresis and Western blotting were performed as previously described . After transfer, the membrane was incubated with TBS buffer supplemented with 0.1% Tween 20 (TBS-T) and 5% BSA at room temperature for 30 min on a shaker, then probed with primary antibodies (pSTAT3 (Cell Signaling D3A7), anti-anti-STAT3 (Cell Signaling 124H6) diluted 1:1000 in TBS buffer supplemented with 0.1% Tween 20 and 3% BSA at 4 °C overnight.…”

Section: Methodsmentioning

confidence: 99%

“…Gel electrophoresis and Western blotting were performed as previously described. 21 After transfer, the membrane was incubated with TBS buffer supplemented with 0.1% Tween 20 (TBS-T) and 5% BSA at room temperature for 30 min on a shaker, then probed with primary antibodies (pSTAT3 (Cell Signaling D3A7), anti-anti-STAT3 (Cell Signaling 124H6) diluted 1:1000 in TBS buffer supplemented with 0.1% Tween 20 and 3% BSA at 4 °C overnight. The membrane was washed with TBS-T four times and probed with secondary antibody (goat antirabbit IgG, DyLight 800 (invitrogen SA535571) IRDye 680RD Goat anti-Mouse IgG (LiCor 926-68070)) at room temperature for 1 h. After another four washes with TBS- T, protein bands on the membrane were visualized using Li-Cor Odyssey CLx and analyzed using StudioLite software.…”

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confidence: 99%

A Small Contribution to a Large System: The Leptin Receptor Complex

et al. 2023

Self Cite

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Obesity is a classified epidemic, increasing the risk of secondary diseases such as diabetes, inflammation, cardiovascular disease, and cancer. The pleiotropic hormone leptin is the proposed link for the gutbrain axis controlling nutritional status and energy expenditure. Research into leptin signaling provides great promise toward discovering therapeutics for obesity and its related diseases targeting leptin and its cognate leptin receptor (LEP-R). The molecular basis underlying the human leptin receptor complex assembly remains obscure, due to the lack of structural information regarding the biologically active complex. In this work, we investigate the proposed receptor binding sites in human leptin utilizing designed antagonist proteins combined with AlphaFold predictions. Our results show that binding site I has a more intricate role in the active signaling complex than previously described. We hypothesize that the hydrophobic patch in this region engages a third receptor forming a higher-order complex, or a new LEP-R binding site inducing allosteric rearrangement.

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“…Purification of 6xHis PLCβ4 followed previously described protocols ( 46 ). Briefly, High Five insect cells were infected with baculovirus at a density between 1.5 × 10 6 and 2 × 10 6 cells/ml at a multiplicity of infection of 1.…”

Section: Methodsmentioning

confidence: 99%

“…Gel electrophoresis and Western blotting were performed as previously described ( 46 ). In brief, after transfer, the membrane was incubated with Tris-buffered saline buffer supplemented with 0.1% Tween-20 (TBS-T) and 5% BSA at room temperature for 30 min on a shaker, then probed with primary antibodies (anti-PLCβ4 [Sigma–Aldrich; catalog no.…”

Section: Methodsmentioning

confidence: 99%