An Unusual U2AF2 Inhibits Splicing and Attenuates the Virulence of the Human Protozoan Parasite Entamoeba histolytica

Abstract

E. histolytica is the etiological agent of intestinal amebiasis and liver abscesses, which still poses public health threat globally. Metronidazole is the drug of choice against amebiasis. However, metronidazole-resistant amoebic clinical isolates and strains have been reported recently, challenging the efforts for amebiasis eradication. In search of alternative treatments, E. histolytica transcriptomes have shown the association of genes involved in RNA metabolism with the virulence of the parasite. Among the upregulated genes in amoebic liver abscesses are the splicing factors EhU2AF2 and a paralog of EhSF3B1. For this reason and because EhU2AF2 contains unusual KH-QUA2 (84KQ) motifs in its lengthened C-terminus domain, here we investigated how the role of EhU2AF2 in pre-mRNA processing impacts the virulence of the parasite. We found that 84KQ is involved in splicing inhibition/intron retention of several virulence and non-virulence-related genes. The 84KQ domain interacts with the same domain of the constitutive splicing factor SF1 (SF1KQ), both in solution and when SF1KQ is bound to branchpoint signal RNA probes. The 84KQ–SF1KQ interaction prevents splicing complex E to A transition, thus inhibiting splicing. Surprisingly, the deletion of the 84KQ domain in EhU2AF2 amoeba transformants increased splicing and enhanced the in vitro and in vivo virulence phenotypes. We conclude that the interaction of the 84KQ and SF1KQ domains, probably involving additional factors, tunes down Entamoeba virulence by favoring intron retention.

The splicing factor SR2 is an important virulence factor of Toxoplasma gondii

“…Splicing factors have been shown to mediate the virulence of various pathogens. For example, KH-QUA2 (84KQ) motifs at the C-terminus of Entamoeba histolytica U2AF2 play roles in splicing of several virulence genes, and absence of 84KQ motifs alters splicing and reduces the parasite virulence (González-Blanco et al, 2022). The deletion of Ustilago maydis Rrm75, which encodes three RRM domains and glycine-rich repeats, fails to produce splicing variants at the 3′-alternative splicing locus of the third exon through AS, resulting in abnormal growth and virulence phenotypes (Rodríguez-Kessler et al, 2012).…”