RESUMEN
The roles of virus-derived small RNAs (vsRNAs) have been studied in plants and insects. However, the generation and function of small RNAs from cytoplasmic RNA viruses in mammalian cells remain unexplored. This study describes four vsRNAs that were detected in enterovirus 71-infected cells using next-generation sequencing and northern blots. Viral infection produced substantial levels (>10(5) copy numbers per cell) of vsRNA1, one of the four vsRNAs. We also demonstrated that Dicer is involved in vsRNA1 generation in infected cells. vsRNA1 overexpression inhibited viral translation and internal ribosomal entry site (IRES) activity in infected cells. Conversely, blocking vsRNA1 enhanced viral yield and viral protein synthesis. We also present evidence that vsRNA1 targets stem-loop II of the viral 5' untranslated region and inhibits the activity of the IRES through this sequence-specific targeting. Our study demonstrates the ability of a cytoplasmic RNA virus to generate functional vsRNA in mammalian cells. In addition, we also demonstrate a potential novel mechanism for a positive-stranded RNA virus to regulate viral translation: generating a vsRNA that targets the IRES.
Asunto(s)
Regiones no Traducidas 5' , Enterovirus Humano A/genética , Regulación Viral de la Expresión Génica , Biosíntesis de Proteínas , ARN Pequeño no Traducido/metabolismo , ARN Viral/metabolismo , Línea Celular Tumoral , Regulación hacia Abajo , Humanos , Ribonucleasa III/metabolismo , Proteínas Virales/biosíntesisRESUMEN
Far-upstream element-binding protein 2 (FBP2) is an internal ribosomal entry site (IRES) trans-acting factor (ITAF) that negatively regulates enterovirus 71 (EV71) translation. This study shows that EV71 infection cleaved FBP2. Live EV71 and the EV71 replicon (but not UV-inactivated virus particles) induced FBP2 cleavage, suggesting that viral replication results in FBP2 cleavage. The results also showed that virus-induced proteasome, autophagy, and caspase activity co-contribute to EV71-induced FBP2 cleavage. Using FLAG-fused FBP2, we mapped the potential cleavage fragments of FBP2 in infected cells. We also found that FBP2 altered its function when its carboxyl terminus was cleaved. This study presents a mechanism for virus-induced cellular events to cleave a negative regulator for viral IRES-driven translation.
Asunto(s)
Enterovirus Humano A/metabolismo , Infecciones por Enterovirus/metabolismo , Biosíntesis de Proteínas/fisiología , Proteínas de Unión al ARN/metabolismo , Transactivadores/metabolismo , Internalización del Virus , Western Blotting , Línea Celular Tumoral , Humanos , Plásmidos/genética , Complejo de la Endopetidasa Proteasomal/metabolismo , ARN Interferente Pequeño/genética , Replicación Viral/fisiologíaRESUMEN
Picornaviruses cause several diseases, not only in humans but also in various animal hosts. For instance, human enteroviruses can cause hand-foot-and-mouth disease, herpangina, myocarditis, acute flaccid paralysis, acute hemorrhagic conjunctivitis, severe neurological complications, including brainstem encephalitis, meningitis and poliomyelitis, and even death. The interaction between the virus and the host is important for viral replication, virulence and pathogenicity. This article reviews studies of the functions of viral and host factors that are involved in the life cycle of picornavirus. The interactions of viral capsid proteins with host cell receptors is discussed first, and the mechanisms by which the viral and host cell factors are involved in viral replication, viral translation and the switch from translation to RNA replication are then addressed. Understanding how cellular proteins interact with viral RNA or viral proteins, as well as the roles of each in viral infection, will provide insights for the design of novel antiviral agents based on these interactions.
Asunto(s)
Infecciones por Picornaviridae/virología , Picornaviridae/fisiología , Animales , Cápside/química , Genoma Viral , Humanos , Modelos Biológicos , Picornaviridae/genética , Infecciones por Picornaviridae/metabolismo , Biosíntesis de Proteínas , Interferencia de ARN , ARN Viral/química , Proteínas Virales/química , Replicación ViralRESUMEN
Enterovirus 71 (EV71) is a neurotropic pathogen that can cause severe neural diseases and complications on infected patients. Clinical observations showed that EV71-induced immune responses may be associated with virus induced neurogenic pulmonary edema. Here reviewed studies that discovered several host molecules as potential factors for EV71 virulence.