The type-1 ribosome-inactivating protein OsRIP1 triggers caspase-independent apoptotic-like death in HeLa cells. (November 2021)

• Record Type: Journal Article
• Authors: Chen, Simin; Lóssio, Cláudia Figueiredo; Verbeke, Isabel; Verduijn, Joost; Parakhonskiy, Bogdan; Van der Meeren, Louis; Chen, Pengyu; De Zaeytijd, Jeroen; Skirtach, André G.; Van Damme, Els J.M.
• Is Part Of: Food and chemical toxicology. Volume 157(2021)
• Publication Date: 2021-11
• Subjects: Ribosome-inactivating protein -- Anti-cancer -- Caspase independent apoptosis -- Cell blebbing -- Cell death; RIPs Ribosome-inactivating proteins -- ASPP1 Apoptosis-Stimulating Protein of p53 1 -- IFITM3 Interferon-induced transmembrane protein 3 -- PUMA P53 upregulated modulator of apoptosis -- IRF1 Interferon regulatory factor 1 -- NHDF normal human dermal fibroblasts -- DMEM Dulbecco's Modifed Eagle's Medium -- BSA Bovine serum albumin -- AFM Atomic force microscopy -- SEM Scanning electron microscopy -- PBS Phosphate-buffered saline -- PFA Paraformaldehyde -- FITC Fluorescein isothiocyanate -- RT-qPCR Quantitative real-time PCR -- GSDMD Gasdermin D -- LC3 microtubule-associated proteins 1A/1B light chain 3B -- MLKL Mixed lineage kinase domain like -- CASP Caspase -- Cyt c Cytochrome c -- IFN Interferon; Toxicology -- Periodicals; Food poisoning -- Periodicals; Food Poisoning -- Periodicals; Toxicology -- Periodicals; Toxicologie -- Périodiques; Intoxications alimentaires -- Périodiques; Food poisoning; Toxicology; Periodicals; Electronic journals; 615.9
• DOI: 10.1016/j.fct.2021.112590
• Languages: English
• ISSNs: 0278-6915

Abstract

Abstract: Ribosome-inactivating proteins (RIPs) are capable of removing a specific adenine from 28S ribosomal RNA, thus inhibiting protein biosynthesis in an irreversible manner. In this study, recombinant OsRIP1, a type 1 RIP from rice ( Oryza sativa L.), was investigated for its anti-proliferative properties. Human cervical cancer HeLa cells were incubated in the presence of OsRIP1 for 24–72 h. OsRIP1 treatment yielded an anti-proliferation response of the HeLa cells and resulted in apoptotic-like blebbing of the plasma membrane without causing DNA fragmentation. OsRIP1 labeled with FITC accumulated at the cell surface. Pull-down assays identified ASPP1 (Apoptosis-Stimulating Protein of p53 1) and IFITM3 (interferon-induced transmembrane protein 3) as potential interaction partners for OsRIP1. Transcript levels for several critical genes related to different signaling pathways were quantified by RT-qPCR. OsRIP1 provoked HeLa cells to undergo caspase-independent cell death, associated with a significant transcriptional upregulation of the apoptotic gene PUMA, interferon regulatory factor 1 (IRF1) and the autophagy-related marker LC3. No changes in caspase activities were observed. Together, these data suggest that apoptotic-like events were involved in OsRIP1-driven caspase-independent cell death that might trigger the IRF1 signaling pathway and LC3-mediated autophagy. Graphical abstract: Image 1 Highlights: Cytotoxicity of the type 1 ribosome inactivating protein OsRIP1 was investigated. OsRIP1 exposure sensitized HeLa cells towards caspase-independent cell death. FITC labeled OsRIP1 accumulated at the cell membrane. Potential interaction partners for OsRIP1 were examined using pull down assays.