Inhibition of Marburg Virus RNA Synthesis by a Synthetic Anti-VP35 Antibody

Overview

Journal ACS Infect Dis

Specialties Infectious Diseases
Microbiology
Pharmacology

Date 2019 May 24

PMID 31120240

Citations 10

Authors

Parmeshwar Amatya

Nicole Wagner

Gang Chen

Priya Luthra

Liuqing Shi

Dominika Borek

Alevtina Pavlenco

Henry Rohrs

Christopher F Basler

Sachdev S Sidhu

Michael L Gross

Daisy W Leung

Affiliations

Soon will be listed here.

Abstract

Marburg virus causes sporadic outbreaks of severe hemorrhagic fever with high case fatality rates. Approved, effective, and safe therapeutic or prophylactic countermeasures are lacking. To address this, we used phage display to engineer a synthetic antibody, sFab H3, which binds the Marburg virus VP35 protein (mVP35). mVP35 is a critical cofactor of the viral replication complex and a viral immune antagonist. sFab H3 displayed high specificity for mVP35 and not for the closely related Ebola virus VP35. sFab H3 inhibited viral-RNA synthesis in a minigenome assay, suggesting its potential use as an antiviral. We characterized sFab H3 by a combination of biophysical and biochemical methods, and a crystal structure of the complex solved to 1.7 Å resolution defined the molecular interface between the sFab H3 and mVP35 interferon inhibitory domain. Our study identifies mVP35 as a therapeutic target using an approach that provides a framework for generating engineered Fabs targeting other viral proteins.

Citing Articles

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References

Fellouse F, Wiesmann C, Sidhu S . Synthetic antibodies from a four-amino-acid code: a dominant role for tyrosine in antigen recognition. Proc Natl Acad Sci U S A. 2004; 101(34):12467-72. PMC: 515084. DOI: 10.1073/pnas.0401786101. View

. The CCP4 suite: programs for protein crystallography. Acta Crystallogr D Biol Crystallogr. 1994; 50(Pt 5):760-3. DOI: 10.1107/S0907444994003112. View

Persson H, Ye W, Wernimont A, Adams J, Koide A, Koide S . CDR-H3 diversity is not required for antigen recognition by synthetic antibodies. J Mol Biol. 2012; 425(4):803-11. PMC: 3764595. DOI: 10.1016/j.jmb.2012.11.037. View

Davis I, Murray L, Richardson J, Richardson D . MOLPROBITY: structure validation and all-atom contact analysis for nucleic acids and their complexes. Nucleic Acids Res. 2004; 32(Web Server issue):W615-9. PMC: 441536. DOI: 10.1093/nar/gkh398. View

Zhu T, Song H, Peng R, Shi Y, Qi J, Gao G . Crystal Structure of the Marburg Virus Nucleoprotein Core Domain Chaperoned by a VP35 Peptide Reveals a Conserved Drug Target for Filovirus. J Virol. 2017; 91(18). PMC: 5571254. DOI: 10.1128/JVI.00996-17. View

Reshetnyak A, Nelson B, Shi X, Boggon T, Pavlenco A, Mandel-Bausch E . Structural basis for KIT receptor tyrosine kinase inhibition by antibodies targeting the D4 membrane-proximal region. Proc Natl Acad Sci U S A. 2013; 110(44):17832-7. PMC: 3816449. DOI: 10.1073/pnas.1317118110. View

Messaoudi I, Amarasinghe G, Basler C . Filovirus pathogenesis and immune evasion: insights from Ebola virus and Marburg virus. Nat Rev Microbiol. 2015; 13(11):663-76. PMC: 5201123. DOI: 10.1038/nrmicro3524. View

Xu H, Freitas M . MassMatrix: a database search program for rapid characterization of proteins and peptides from tandem mass spectrometry data. Proteomics. 2009; 9(6):1548-55. PMC: 2759086. DOI: 10.1002/pmic.200700322. View

Luthra P, Naidoo J, Pietzsch C, De S, Khadka S, Anantpadma M . Inhibiting pyrimidine biosynthesis impairs Ebola virus replication through depletion of nucleoside pools and activation of innate immune responses. Antiviral Res. 2018; 158:288-302. PMC: 6436837. DOI: 10.1016/j.antiviral.2018.08.012. View

10.

Zemlin M, Klinger M, Link J, Zemlin C, Bauer K, Engler J . Expressed murine and human CDR-H3 intervals of equal length exhibit distinct repertoires that differ in their amino acid composition and predicted range of structures. J Mol Biol. 2003; 334(4):733-49. DOI: 10.1016/j.jmb.2003.10.007. View

11.

Kirchdoerfer R, Abelson D, Li S, Wood M, Saphire E . Assembly of the Ebola Virus Nucleoprotein from a Chaperoned VP35 Complex. Cell Rep. 2015; 12(1):140-149. PMC: 4500542. DOI: 10.1016/j.celrep.2015.06.003. View

12.

Luthra P, Liang J, Pietzsch C, Khadka S, Edwards M, Wei S . A high throughput screen identifies benzoquinoline compounds as inhibitors of Ebola virus replication. Antiviral Res. 2018; 150():193-201. PMC: 11727378. DOI: 10.1016/j.antiviral.2017.12.019. View

13.

Liu G, Nash P, Johnson B, Pietzsch C, Ilagan M, Bukreyev A . A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35-NP Interface. ACS Infect Dis. 2017; 3(3):190-198. PMC: 5735849. DOI: 10.1021/acsinfecdis.6b00209. View

14.

Muhlberger E, Weik M, Volchkov V, Klenk H, Becker S . Comparison of the transcription and replication strategies of marburg virus and Ebola virus by using artificial replication systems. J Virol. 1999; 73(3):2333-42. PMC: 104478. DOI: 10.1128/JVI.73.3.2333-2342.1999. View

15.

Leung D, Ginder N, Fulton D, Nix J, Basler C, Honzatko R . Structure of the Ebola VP35 interferon inhibitory domain. Proc Natl Acad Sci U S A. 2009; 106(2):411-6. PMC: 2626716. DOI: 10.1073/pnas.0807854106. View

16.

Xu W, Luthra P, Wu C, Batra J, Leung D, Basler C . Ebola virus VP30 and nucleoprotein interactions modulate viral RNA synthesis. Nat Commun. 2017; 8:15576. PMC: 5472179. DOI: 10.1038/ncomms15576. View

17.

Seesuay W, Jittavisutthikul S, Sae-Lim N, Sookrung N, Sakolvaree Y, Chaicumpa W . Human transbodies that interfere with the functions of Ebola virus VP35 protein in genome replication and transcription and innate immune antagonism. Emerg Microbes Infect. 2018; 7(1):41. PMC: 5864874. DOI: 10.1038/s41426-018-0031-3. View

18.

Bruhn J, Kirchdoerfer R, Urata S, Li S, Tickle I, Bricogne G . Crystal Structure of the Marburg Virus VP35 Oligomerization Domain. J Virol. 2016; 91(2). PMC: 5215338. DOI: 10.1128/JVI.01085-16. View

19.

Miersch S, Sidhu S . Intracellular targeting with engineered proteins. F1000Res. 2016; 5. PMC: 4984483. DOI: 10.12688/f1000research.8915.1. View

20.

Muhlberger E, Lotfering B, Klenk H, Becker S . Three of the four nucleocapsid proteins of Marburg virus, NP, VP35, and L, are sufficient to mediate replication and transcription of Marburg virus-specific monocistronic minigenomes. J Virol. 1998; 72(11):8756-64. PMC: 110291. DOI: 10.1128/JVI.72.11.8756-8764.1998. View