Redox-sensitive small GTPase H-Ras in murine astrocytes, an in vitro study

Redox Rep. 2022 Dec;27(1):150-157. doi: 10.1080/13510002.2022.2094109.

Abstract

Background: Although the protooncogenes small GTPases Ras are redox-sensitive proteins, how they are regulated by redox signaling in the central nervous system (CNS) is still poorly understood. Alteration in redox-sensitive targets by redox signaling may have myriad effects on Ras stability, activity and localization. Redox-mediated changes in astrocytic RAS may contribute to the control of redox homeostasis in the CNS that is connected to the pathogenesis of many diseases.

Results and methods: Here, we investigated the transient physiological induction, at both transcriptional and translational levels, of small GTPases Ras in response to redox stimulation. Cultured astrocytes were treated with hydrogen peroxide as in bolus addition and relative mRNA levels of murine hras and kras genes were detected by qRT-PCR. We found that de novo transcription of hras mRNA in reactive astrocytes is redox-sensitive and mimics the prototypical redox-sensitive gene iNOS. Protein abundance in combination with protein turnover measurements by cycloheximide-chase experiments revealed distinct translation efficiency, GTP-bound enrichment, and protein turnover rates between the two isoforms H-Ras and K-Ras.

Conclusion: Reports from recent years support a significant role of H-Ras in driving redox processes. Beyond its canonical functions, Ras may impact on the core astrocytic cellular machinery that operates during redox stimulation.

Keywords: GFAP; Small GTPase Ras; murine astrocytes; redox signaling.

MeSH terms

  • Animals
  • Astrocytes / metabolism
  • Mice
  • Monomeric GTP-Binding Proteins* / metabolism
  • Oxidation-Reduction
  • Proto-Oncogene Proteins p21(ras)* / metabolism
  • RNA, Messenger / metabolism
  • ras Proteins / genetics
  • ras Proteins / metabolism

Substances

  • RNA, Messenger
  • Hras protein, mouse
  • Monomeric GTP-Binding Proteins
  • Proto-Oncogene Proteins p21(ras)
  • ras Proteins

Grants and funding

The author(s) reported there is no funding associated with the work featured in this article.