Synonymous Changes in Specific Leucine Codons Impact Morbilliviral Protein Production from Human & Canine Codon Optimized Constructs

Abstract

Protein expression is increased by optimization of human measles virus (HMV) & canine distemper virus (CDV) nucleoprotein (N) gene to human or canine codon usage bias (CUB) and decreased by suboptimization. Six leucine codons were all replaced by CTG in human CUB optimized N gene constructs and by CTG & CTC in canine CUB optimized N gene constructs. For CUB suboptimized N gene constructs only CTT coded for leucine. The purpose of this experiment was to determine the effects of synonymous leucine codon changes. Protein expression from human & canine CUB optimized constructs in which CTG was replaced with CTT was compared using flow cytometry to that from WT, human & canine CUB optimized versions. Protein expression from CUB suboptimized versions with CTT changed to CTG was also compared to that from unmodified constructs. Human & canine CUB optimized CTT versions of the HMV N gene had significantly decreased protein production, with expression from the CTT canine CUB optimized construct the most suppressed. Replacing CTG with CTT in the human optimized CDV N gene constructs decreased the level of protein expression, while changing CTG to CTT in the canine optimized CDV N gene construct did not significantly change the level of protein expression. Protein expression from the CTT modified suboptimized CDV N gene construct was significantly increased compared to that from the unmodified suboptimized version. These results indicate that synonymous changes in specific leucine codons can significantly impact morbilliviral protein production in ways that vary with the virus and the context in which they occur.Support or Funding InformationThe project depicted was sponsored by the Department of Defense Threat Reduction Agency. The content of the information does not necessarily reflect the position or the policy of the federal government, and no official endorsement should be inferredThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.