Viral RNA-binding Proteins Research Articles

Patterns for epilepsy care

<h3>ABSTRACT</h3> Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes COVID-19. SARS-CoV-2 relies on cellular RNA-binding proteins (RBPs) to replicate and spread, although which RBPs control SARS-CoV-2 infection remains largely unknown. Here, we employ a multi-omic approach to identify systematically and comprehensively which cellular and viral RBPs are involved in SARS-CoV-2 infection. We reveal that the cellular RNA-bound proteome is remodelled upon SARS-CoV-2 infection, having widespread effects on RNA metabolic pathways, non-canonical RBPs and antiviral factors. Moreover, we apply a new method to identify the proteins that directly interact with viral RNA, uncovering dozens of cellular RBPs and six viral proteins. Amongst them, several components of the tRNA ligase complex, which we show regulate SARS-CoV-2 infection. Furthermore, we discover that available drugs targeting host RBPs that interact with SARS-CoV-2 RNA inhibit infection. Collectively, our results uncover a new universe of host-virus interactions with potential for new antiviral therapies against COVID-19.

Translational Control by Influenza Virus: IDENTIFICATION OF CIS-ACTING SEQUENCES AND TRANS-ACTING FACTORS WHICH MAY REGULATE SELECTIVE VIRAL mRNA TRANSLATION

We have shown that sequences contained within the viral mRNA 5'-untranslated region (UTR) played a critical role in directing selective influenza viral mRNA translation. We therefore attempted to identify transacting factors that may regulate viral mRNA translation through interactions with the 5'-UTR and at the same time map the precise sequences to which these factors bind. We can now demonstrate that multiple cellular proteins interact with influenza viral but not cellular 5'-UTRs using gel mobility shift and UV cross-linking analyses. Gel supershift studies revealed that the La autoantigen was one of the cellular proteins that interacted with the viral 5'-UTR. Utilizing mutants of the viral mRNA 5' UTR, we have determined that sequences within the very 5'-conserved region and nucleotides immediately 3' are necessary but not always sufficient for binding certain cellular proteins. Northwestern analysis showed the binding of a distinct subset of cellular proteins to the viral 5'-UTR, but also demonstrated interactions of the viral nonstructural protein NS1. Gel shift analysis with purified recombinant NS1 confirmed the binding of the viral protein to a specific region of the viral 5'-UTRs. A model describing the possible role of these cellular and viral RNA-binding proteins in regulating influenza virus mRNA translation will be discussed.

RNA-binding proteins of coronavirus MHV: Detection of monomeric and multimeric N protein with an RNA overlay-protein blot assay

RNA-binding proteins of coronavirus MHV-A59 were identified using an RNA overlay-protein blot assay (ROPBA). The major viral RNA-binding protein in virions and infected cells was the phosphorylated nucleocapsid protein N (50K). A new 140K virus structural protein was identified as a minor RNA-binding protein both in virions and in infected cells. The 140K protein was antigenically related to N, and upon reduction, yielded only 50K N. Thus, the 140K protein is probably a trimer of N subunits linked by intermolecular disulfide bonds. Several cellular RNA-binding proteins were also detected. RNA-binding of N was not nucleotide sequence specific. Single-stranded RNA of MHV, VSV, or cellular origin, a DNA probe of the MHV leader sequence, and double-stranded bovine rotavirus RNA could all bind to N. Binding of MHV RNA was optimal between pH 7 and 8, and the RNA could be eluted in 0.1 M NaCl. The ROPBA is a useful method for the initial identification of RNA-binding proteins, such as N and the 140K protein of murine coronavirus.