5D8U

2009 H1N1 PA endonuclease mutant E119D in complex with L-742,001

Summary for 5D8U

• Entry DOI: 10.2210/pdb5d8u/pdb
• Related: 5CCY 5CGV 5CL0 5CZN 5D2O 5D42 5D4G 5D9J
• Descriptor: Polymerase acidic protein, MANGANESE (II) ION, (2Z)-4-[1-benzyl-4-(4-chlorobenzyl)piperidin-4-yl]-2-hydroxy-4-oxobut-2-enoic acid, ... (4 entities in total)
• Functional Keywords: influenza, resistance, endonuclease inhibitor, viral protein, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• Biological source: Influenza A virus; More
• Total number of polymer chains: 1
• Total formula weight: 23658.09

Authors

Kumar, G.,White, S.W. (deposition date: 2015-08-18, release date: 2016-03-16, Last modification date: 2023-09-27)

Primary citation

Song, M.S.,Kumar, G.,Shadrick, W.R.,Zhou, W.,Jeevan, T.,Li, Z.,Slavish, P.J.,Fabrizio, T.P.,Yoon, S.W.,Webb, T.R.,Webby, R.J.,White, S.W.
Identification and characterization of influenza variants resistant to a viral endonuclease inhibitor.
Proc.Natl.Acad.Sci.USA, 113:3669-3674, 2016

PubMed Abstract: The influenza endonuclease is an essential subdomain of the viral RNA polymerase. It processes host pre-mRNAs to serve as primers for viral mRNA and is an attractive target for antiinfluenza drug discovery. Compound L-742,001 is a prototypical endonuclease inhibitor, and we found that repeated passaging of influenza virus in the presence of this drug did not lead to the development of resistant mutant strains. Reduced sensitivity to L-742,001 could only be induced by creating point mutations via a random mutagenesis strategy. These mutations mapped to the endonuclease active site where they can directly impact inhibitor binding. Engineered viruses containing the mutations showed resistance to L-742,001 both in vitro and in vivo, with only a modest reduction in fitness. Introduction of the mutations into a second virus also increased its resistance to the inhibitor. Using the isolated wild-type and mutant endonuclease domains, we used kinetics, inhibitor binding and crystallography to characterize how the two most significant mutations elicit resistance to L-742,001. These studies lay the foundation for the development of a new class of influenza therapeutics with reduced potential for the development of clinical endonuclease inhibitor-resistant influenza strains.

PubMed: 26976575
DOI: 10.1073/pnas.1519772113

Experimental method

X-RAY DIFFRACTION (2.29 Å)