Chikungunya virus (CHIKV) infection induces the formation of migrasomes, yet their specific role in CHIKV pathogenesis remains unclear. This study explores the mechanisms underlying mitochondrial damage induced by CHIKV 181 clone 25 (CHIKV 181/25) and the role of migrasomes in mitigating this damage. Using cultured cell lines, we assessed the impact of CHIKV infection on mitochondrial integrity and function, with particular emphasis on the viroporin proteins transframe (TF) and 6K. We utilized fluorescence microscopy and transmission electron microscopy to visualize the interplay between migrasome formation and damaged mitochondria. Additionally, calcium imaging assays were conducted to evaluate intracellular calcium levels, and RNA sequencing was performed to examine gene expression. Our results demonstrated that CHIKV infection leads to mitochondrial damage, mediated by the action of TF and 6K. Notably, migrasomes induced by nonstructural protein 1 (nsP1) effectively clearing impaired mitochondria through mitocytosis. Furthermore, we identified the arginine residue R37 within the viroporin proteins of CHIKV as crucial for inducing mitochondrial damage through elevated intracellular calcium levels. Importantly, R37 within TF from other alphaviruses is also critical for mitochondrial damage. In conclusion, our findings elucidate the complex interplay between CHIKV and mitochondrial dysfunction, positioning migrasomes as potential mediators in alleviating CHIKV-induced mitochondrial damage.

O-glycosylation is an ancient yet underappreciated protein posttranslational modification, on which many bacteria and viruses heavily rely to perform critical biological functions involved in numerous infectious diseases or even cancer. But due to the innate complexity of O-glycosylation, research techniques have been limited to study its exact role in viral attachment and entry, assembly and exit, spreading in the host cells, and the innate and adaptive immunity of the host. Recently, the advent of many newly developed methodologies (e.g., mass spectrometry, chemical biology tools, and molecular dynamics simulations) has renewed and rekindled the interest in viral-related O-glycosylation in both viral proteins and host cells, which is further fueled by the COVID-19 pandemic. In this review, we summarize recent advances in viral-related O-glycosylation, with a particular emphasis on the mucin-type O-linked α-N-acetylgalactosamine (O-GalNAc) on viral proteins and the intracellular O-linked β-N-acetylglucosamine (O-GlcNAc) modifications on host proteins. We hope to provide valuable insights into the development of antiviral reagents or vaccines for better prevention or treatment of infectious diseases.