Two Domains That Control Prefusion Stability and Transport Competence of the Measles Virus Fusion Protein

Doyle, Joshua; Prussia, Andrew; White, Laura K.; Sun, Aiming; Liotta, Dennis; Snyder, James P.; Compans, Richard W.; Plemper, Richard K.

doi:10.1128/jvi.80.3.1524-1536.2006

Cited by 49 publications

(91 citation statements)

References 60 publications

(90 reference statements)

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“…Moreover, resistance to the compound (AS-48) coincided with increased fusion activity (45,46). We now extend this notion by demonstrating that resistance mutations locating in a "pocket" microdomain within the globular head domain reduce the stability of prefusion F trimers.…”

supporting

confidence: 54%

“…We propose that this F mutant is very likely impaired by one or multiple additional steps required for productive fusion pore formation. drug, and several resistant F variants showed a hyperfusogenic phenotype (45,46). These results strongly suggested that mutant F trimers resisted the AS-48 compound by a mechanism other than prevention of direct docking of the molecule.…”

Section: Resultsmentioning

confidence: 87%

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Molecular Determinants Defining the Triggering Range of Prefusion F Complexes of Canine Distemper Virus

Avila

Alves

Khosravi

et al. 2014

J Virol

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The morbillivirus cell entry machinery consists of a fusion (F) protein trimer that refolds to mediate membrane fusion following receptor-induced conformational changes in its binding partner, the tetrameric attachment (H) protein. To identify molecular determinants that control F refolding, we generated F chimeras between measles virus (MeV) and canine distemper virus (CDV). We located a central pocket in the globular head domain of CDV F that regulates the stability of the metastable, prefusion conformational state of the F trimer. Most mutations introduced into this "pocket'" appeared to mediate a destabilizing effect, a phenotype associated with enhanced membrane fusion activity. Strikingly, under specific triggering conditions (i.e., variation of receptor type and H protein origin), some F mutants also exhibited resistance to a potent morbillivirus entry inhibitor, which is known to block F triggering by enhancing the stability of prefusion F trimers. Our data reveal that the molecular nature of the F stimulus and the intrinsic stability of metastable prefusion F both regulate the efficiency of F refolding and escape from smallmolecule refolding blockers. IMPORTANCEWith the aim to better characterize the thermodynamic basis of morbillivirus membrane fusion for cell entry and spread, we report here that the activation energy barrier of prefusion F trimers together with the molecular nature of the triggering "stimulus" (attachment protein and receptor types) define a "triggering range," which governs the initiation of the membrane fusion process. A central "pocket" microdomain in the globular F head contributes substantially to the regulation of the conformational stability of the prefusion complexes. The triggering range also defines the mechanism of viral escape from entry inhibitors and describes how the cellular environment can affect membrane fusion efficiency.

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supporting

confidence: 54%

Section: Resultsmentioning

confidence: 87%

Molecular Determinants Defining the Triggering Range of Prefusion F Complexes of Canine Distemper Virus

Avila

Alves

Khosravi

et al. 2014

J Virol

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“…We demonstrate that the 400 and 489 microdomains are located at opposing ends of the postfusion F structure, but are posited in close proximity to each other at the intersection of stalk and head domain in prefusion F. Interestingly, several studies investigating related paramyxovirus F proteins have identified this network of noncovalent interactions between prefusion F stalk and head as a major determinant for controlling the conformational stability of the trimer (49,50). We have furthermore demonstrated that point mutations in this region confer resistance against a small-molecule entry inhibitor of MeV that we have developed (51). Second, point mutations in either of the two resistance hotspots reduced the structural stability of the prefusion RSV F trimer in biochemical assays and resulted in enhanced spontaneous viral inactivation rates in the absence of target cells.…”

Section: Discussionmentioning

confidence: 72%

Cross-resistance mechanism of respiratory syncytial virus against structurally diverse entry inhibitors

Yan

Lee

Thakkar

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

107

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Respiratory syncytial virus (RSV) is a leading pediatric pathogen that is responsible for a majority of infant hospitalizations due to viral disease. Despite its clinical importance, no vaccine prophylaxis against RSV disease or effective antiviral therapeutic is available. In this study, we established a robust high-throughput drug screening protocol by using a recombinant RSV reporter virus to expand the pool of RSV inhibitor candidates. Mechanistic characterization revealed that a potent newly identified inhibitor class blocks viral entry through specific targeting of the RSV fusion (F) protein. Resistance against this class was induced and revealed overlapping hotspots with diverse, previously identified RSV entry blockers at different stages of preclinical and clinical development. A structural and biochemical assessment of the mechanism of unique, broad RSV cross-resistance against structurally distinct entry inhibitors demonstrated that individual escape hotspots are located in immediate physical proximity in the metastable conformation of RSV F and that the resistance mutations lower the barrier for prefusion F triggering, resulting in an accelerated RSV entry kinetics. One resistant RSV recombinant remained fully pathogenic in a mouse model of RSV infection. By identifying molecular determinants governing the RSV entry machinery, this study spotlights a molecular mechanism of broad RSV resistance against entry inhibition that may affect the impact of diverse viral entry inhibitors presently considered for clinical use and outlines a proactive design for future RSV drug discovery campaigns.antiviral drugs | drug resistance

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“…There was no amino-acid substitution in their H (Htag) and M proteins. The F(N462K) protein has already been reported to exhibit enhanced fusion activity 11 .…”

Section: Fusion Assay Inconsistent With One Recombinant Virusmentioning

confidence: 99%

Cooperation between different RNA virus genomes produces a new phenotype

2012

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Two Domains That Control Prefusion Stability and Transport Competence of the Measles Virus Fusion Protein

Cited by 49 publications

References 60 publications

Molecular Determinants Defining the Triggering Range of Prefusion F Complexes of Canine Distemper Virus

Molecular Determinants Defining the Triggering Range of Prefusion F Complexes of Canine Distemper Virus

Cross-resistance mechanism of respiratory syncytial virus against structurally diverse entry inhibitors

Cooperation between different RNA virus genomes produces a new phenotype

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