Protein Sumoylation Is Crucial for Phagocytosis in <i>Entamoeba histolytica</i> Trophozoites

• Main Authors: Mitzi Díaz-Hernández, Rosario Javier-Reyna, Izaid Sotto-Ortega, Guillermina García-Rivera, Sarita Montaño, Abigail Betanzos, Dxinegueela Zanatta, Esther Orozco
• Format: Article
• Language: English
• Published: MDPI AG 2021-05-01
• Series: International Journal of Molecular Sciences
• Subjects: SUMOylation; phagocytosis; <i>E. histolytica</i>; ESCRT machinery; EhADH adhesin
• Online Access: https://www.mdpi.com/1422-0067/22/11/5709

Posttranslational modifications provide <i>Entamoeba histolytica</i> proteins the timing and signaling to intervene during different processes, such as phagocytosis. However, SUMOylation has not been studied in <i>E. histolytica</i> yet. Here, we characterized the <i>E. histolytica SUMO</i> gene, its product (EhSUMO), and the relevance of SUMOylation in phagocytosis. Our results indicated that EhSUMO has an extended N-terminus that differentiates SUMO from ubiquitin. It also presents the GG residues at the C-terminus and the ΨKXE/D binding motif, both involved in target protein contact. Additionally, the <i>E. histolytica</i> genome possesses the enzymes belonging to the SUMOylation-deSUMOylation machinery. Confocal microscopy assays disclosed a remarkable EhSUMO membrane activity with convoluted and changing structures in trophozoites during erythrophagocytosis. SUMOylated proteins appeared in pseudopodia, phagocytic channels, and around the adhered and ingested erythrocytes. Docking analysis predicted interaction of EhSUMO with EhADH (an ALIX family protein), and immunoprecipitation and immunofluorescence assays revealed that the association increased during phagocytosis; whereas the EhVps32 (a protein of the ESCRT-III complex)-EhSUMO interaction appeared stronger since basal conditions. In <i>EhSUMO</i> knocked-down trophozoites, the bizarre membranous structures disappeared, and EhSUMO interaction with EhADH and EhVps32 diminished. Our results evidenced the presence of a <i>SUMO</i> gene in <i>E. histolytica</i> and the SUMOylation relevance during phagocytosis. This is supported by bioinformatics screening of many other proteins of <i>E. histolytica</i> involved in phagocytosis, which present putative SUMOylation sites and the ΨKXE/D binding motif.