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bioRxiv · 10.1101/2023.05.20.541575

C-terminal cysteines of HRas control Erk signaling and 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) mediated inhibition of myoblast differentiation.

Pundlik, S. S.·Sahoo, S. S.·Barik, A.·Jaysingh, M. A.·Venkateshvaran, A.·Math, R. G. H.·Ramanathan, A.
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Abstract

HRas is an important node that controls cellular signaling, proliferation, and differentiation. Mutants of HRas (e.g., the constitutively active HRas V12) can be oncogenic, and can also inhibit myoblast differentiation. The C-terminal cysteines of HRas (Cys181 and Cys184) serve as substrates for intra-cellular reversible palmitoylation and de-palmitoylation reactions, which control its subcellular distribution. The relationship between the C-terminal cysteines of HRas, its intracellular distribution, and its cellular activity has remained unclear. Understanding this relationship has important implications for targeting HRas in pathogenic states where it is activated. In this study, we show that a mutation in the C-terminal of HRas, C181S, is sufficient to cause increased levels of HRas V12 in the Golgi, decreased HRas V12-driven Akt and Erk signaling and reverse the ability of HRas V12 to inhibit myoblast differentiation. This demonstrates the importance of C-terminal cysteines in controlling HRas V12. It has been previously shown that Cys184 can also be irreversibly modified by an electrophilic prostaglandin lipid 15d-PGJ2. This lipid is released by senescent cells as a part of senescence-associated secretory phenotype (SASP). In this study, we show that 15d-PGJ2 is secreted by senescent myoblasts formed by treatment with Doxorubicin. We also show that 15d-PGJ2 causes decreased levels of HRas within Golgi, activates Erk signaling (but not Akt signaling), and inhibits differentiation of C2C12 myoblasts in an HRas Cys184-dependent fashion. Chemotherapeutics such as Doxorubicin drive senescence and loss of skeletal muscle homeostasis in cancer patients. This study suggests that targeting the senescence-derived synthesis of 15-PGJ2 might be a target to promote muscle homeostasis after chemotherapy.

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Pundlik, S. S., Sahoo, S. S., Barik, A., Jaysingh, M. A., Venkateshvaran, A., Math, R. G. H., Ramanathan, A.. 2023-05-22. C-terminal cysteines of HRas control Erk signaling and 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) mediated inhibition of myoblast differentiation.. https://doi.org/10.1101/2023.05.20.541575

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