Primary resistance mechanism of the canine distemper virus fusion protein against a small-molecule membrane fusion inhibitor.

Kalbermatter, David; Shrestha, Neeta; Ebert, Nadine; Herren, Michael; Moll, Pascal Dominique; Plemper, Richard K; Altmann, Karl-Heinz; Langedijk, Johannes P; Gall, Flavio; Lindenmann, Urs; Riedl, Rainer; Fotiadis, Dimitrios José; Plattet, Philippe (2019). Primary resistance mechanism of the canine distemper virus fusion protein against a small-molecule membrane fusion inhibitor. Virus research, 259, pp. 28-37. Elsevier 10.1016/j.virusres.2018.10.003

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Morbilliviruses (e.g. measles virus [MeV] or canine distemper virus [CDV]) employ the attachment (H) and fusion (F) envelope glycoproteins for cell entry. H protein engagement to a cognate receptor eventually leads to F-triggering. Upon activation, F proteins transit from a prefusion to a postfusion conformation; a refolding process that is associated with membrane merging. Small-molecule morbilliviral fusion inhibitors such as the compound 3G (a chemical analog in the AS-48 class) were previously generated and mechanistic studies revealed a stabilizing effect on morbilliviral prefusion F trimers. Here, we aimed at designing 3G-resistant CDV F mutants by introducing single cysteine residues at hydrophobic core positions of the helical stalk region. Covalently-linked F dimers were generated, which highlighted substantial conformational flexibility within the stalk to achieve those irregular F conformations. Our findings demonstrate that "top-stalk" CDV F cysteine mutants (F-V571C and F-L575C) remained functional and gained resistance to 3G. Conversely, although not all "bottom-stalk" F cysteine variants preserved proper bioactivity, those that remained functional exhibited 3G-sensitivity. According to the recently determined prefusion MeV F trimer/AS-48 co-crystal structure, CDV residues F-V571 and F-L575 may directly interact with 3G. A combination of conformation-specific anti-F antibodies and low-resolution electron microscopy structural analyses confirmed that 3G lost its stabilizing effect on "top-stalk" F cysteine mutants thus suggesting a primary resistance mechanism. Overall, our data suggest that the fusion inhibitor 3G stabilizes prefusion CDV F trimers by docking at the top of the stalk domain.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Faculty Institutions > NCCR TransCure
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Experimental Clinical Research
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH)

UniBE Contributor:

Kalbermatter, David, Shrestha, Neeta, Ebert, Nadine, Herren, Michael, Moll, Pascal Dominique, Fotiadis, Dimitrios José, Plattet, Philippe

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health
600 Technology > 630 Agriculture

ISSN:

0168-1702

Publisher:

Elsevier

Language:

English

Submitter:

Barbara Franziska Järmann-Bangerter

Date Deposited:

14 Feb 2019 08:10

Last Modified:

05 Dec 2022 15:24

Publisher DOI:

10.1016/j.virusres.2018.10.003

PubMed ID:

30296457

Uncontrolled Keywords:

Fusion inhibitors Fusion protein Head-stalk interface Morbillivirus cell entry Prefusion state stabilization

BORIS DOI:

10.7892/boris.124482

URI:

https://boris.unibe.ch/id/eprint/124482

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