Defining the functional determinants for RNA surveillance by RIG-I

By Andrew S. Kohlway, D. Luo, David C. Rawling, S. Ding, Anna Marie Pyle. 2013

111 citations recorded by the source corpus.

Retinoic acid‐inducible gene‐I (RIG‐I) is an intracellular RNA sensor that activates the innate immune machinery in response to infection by RNA viruses. Here, we report the crystal structure of distinct conformations of a RIG‐I:dsRNA complex, which shows that HEL2i‐mediated scanning allows RIG‐I to sense the length of RNA targets. To understand the implications of HEL2i scanning for catalytic activity and signalling by RIG‐I, we examined its ATPase activity when stimulated by duplex RNAs of varying lengths and 5′ composition. We identified a minimal RNA duplex that binds one RIG‐I molecule, stimulates robust ATPase activity, and elicits a RIG‐I‐mediated interferon response in cells. Our results reveal that the minimal functional unit of the RIG‐I:RNA complex is a monomer that binds at the terminus of a duplex RNA substrate. This behaviour is markedly different from the RIG‐I paralog melanoma differentiation‐associated gene 5 (MDA5), which forms cooperative filaments.

Subfields: Immunology, Molecular Biology

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Immunology

In Molecular Biology

Citations in the corpus

The RIG-I receptor adopts two different conformations for distinguishing host from viral RNA ligands (2022, neutral): “, 2018), but it is 16 base pairs longer (see STAR Methods), and it readily stimulates RIG-I activation (Figure 5A; Kohlway et al., 2013).”
Selective RNA targeting and regulated signaling by RIG-I is controlled by coordination of RNA and ATP binding (2016, methodology): “The ATPase activity of S411A is similarly reduced when stimulated by OH10L (data not shown), which was previously shown to activate WT RIG-I ATPase, but not signaling activity (22,41).”
Unified mechanisms for self-RNA recognition by RIG-I Singleton-Merten syndrome variants (2018, unknown): “In accordance with previous data (Kohlway et al., 2013), the presence of RNA increases the affinity of wtRIG-I and RIG-I E373Q, but not of RIG-I K270I for MANT-ATP or MANTATPgS.”
The RIG-I ATPase core has evolved a functional requirement for allosteric stabilization by the Pincer domain (2014, methodology): “Recombinant RIG-I proteins were expressed and purified as reported previously (12,35).”
Kinetic discrimination of self/non-self RNA by the ATPase activity of RIG-I and MDA5 (2015, unknown): “A series of crystal structures shows the hel2i residues K508 and Q511 contacting several different base pairs along the dsRNA, which has been interpreted to reflect a sequential scanning movement of the hel2i domain along a 10-mer dsRNA [19].”
Establishing the role of ATP for the function of the RIG-I innate immune sensor (2015, methodology): “While robust signaling is often associated with ATP hydrolysis activity (Yoneyama et al., 2004; Bamming and Horvath, 2009; Luo et al., 2011; Kohlway et al., 2013; Patel et al., 2013; Peisley et al., 2013; Rawling et al., 2015), the ability of an RNA ligand to stimulate RIG-I ATPase activity does…”
RNA binding activates RIG-I by releasing an autorepressed signaling domain (2019, background): “For example, RNAs that bind RIG-I or stimulate ATP hydrolysis do not always signal in cells (18, 20).”
Influenza A Virus Panhandle Structure Is Directly Involved in RIG-I Activation and Interferon Induction (2015, background): “In addition, because the EMSA only revealed one shifted band of the complex, this interaction most likely reflects a binding scenario with 1:1 stoichiometry, as previously reported for synthetic RNA of similar size (46).”
ATP hydrolysis by the viral RNA sensor RIG-I prevents unintentional recognition of self-RNA (2015, background): “…approximately matches the footprint of dsRNA across the two SF2 RecA domains in the crystal structure of ADP BeFx-bound RIG-I (Jiang et al., 2011; Kohlway et al., 2013; Kowalinski et al., 2011; Luo et al., 2011) and also meets the requirements for activation of RIG-I’s ATPase Figure supplement 1.”
RIG-I uses an ATPase-powered translocation-throttling mechanism for kinetic proofreading of RNAs and oligomerization (2018, background): “Although RIG-I has a lower affinity for nonPAMPs, differences in the binding affinities for PAMP and non-PAMP RNAs (~20–500 fold) are not sufficient to explain the high selectivity of RIG-I activation (Devarkar et al., 2016; Kohlway et al., 2013; Ramanathan et al., 2016).”
Structural basis for m7G recognition and 2′-O-methyl discrimination in capped RNAs by the innate immune receptor RIG-I (2016, unknown): “RNAs carrying a 5 ′ -triphosphate (5 ′ ppp) moiety and blunt-ended double-stranded (ds)RNAs are the best characterized PAMP ligands of RIG-I, showing high-affinity binding and robust stimulation of the ATP hydrolysis activity (8 – 10).”
A minimal RNA ligand for potent RIG-I activation in living mice (2017, background): “Previous work has suggested that RIG-I function can be controlled and exploited pharmacologically through stimulation with small, well-defined RNA ligands that are no larger than other therapeutically administered oligonucleotides (9, 10, 16).”
Sequence-Specific Modifications Enhance the Broad-Spectrum Antiviral Response Activated by RIG-I Agonists (2015, background): “Interestingly, the highly conserved 5= and 3= untranslated regions of negative single-strand RNA virus genomes display high base pair complementarity, and the blunt terminus of the RNA panhandle structure theoretically formed by the viral genome meets the minimal requirements for RIG-I recognition a”