Accurate Measurement of the Effects of All Amino-Acid Mutations on Influenza Hemagglutinin.


Influenza genes evolve mostly via point mutations, and so knowing the effect of every amino-acid mutation provides information about evolutionary paths available to the virus. We and others have combined high-throughput mutagenesis with deep sequencing to estimate the effects of large numbers of mutations to influenza genes. However, these measurements have suffered from substantial experimental noise due to a variety of technical problems, the most prominent of which is bottlenecking during the generation of mutant viruses from plasmids. Here we describe advances that ameliorate these problems, enabling us to measure with greatly improved accuracy and reproducibility the effects of all amino-acid mutations to an H1 influenza hemagglutinin on viral replication in cell culture. The largest improvements come from using a helper virus to reduce bottlenecks when generating viruses from plasmids. Our measurements confirm at much higher resolution the results of previous studies suggesting that antigenic sites on the globular head of hemagglutinin are highly tolerant of mutations. We also show that other regions of hemagglutinin-including the stalk epitopes targeted by broadly neutralizing antibodies-have a much lower inherent capacity to tolerate point mutations. The ability to accurately measure the effects of all influenza mutations should enhance efforts to understand and predict viral evolution.

Submission Details

ID: EepmqWV43

Submitter: Shu-Ching Ou

Submission Date: July 23, 2018, 4:22 p.m.

Version: 1

Publication Details
Doud MB;Bloom JD,Viruses (2016) Accurate Measurement of the Effects of All Amino-Acid Mutations on Influenza Hemagglutinin. PMID:27271655
Additional Information

Number of data points 11280
Proteins Hemagglutinin
Unique complexes 10717
Assays/Quantities/Protocols Experimental Assay: Generation of Mutant HA Virus Libraries
Libraries Overall Merged Site-Specific Amino-Acid Preferences
Sequence Assay Result Units