HCoV-229E N-protein phosphorylation

The aim of the experiment is to identify and compare changes in host gene expression of Huh-7 cells (human liver carcinoma cells) infected with Human coronavirus 229E (HCoV-229E, WT) or genetically engineered HCoV-229E mutants in which the phosphorylation status of the structural N protein has been altered by two combined Ser-to-Ala substitutions in each of the two mutants (S147A+S149A and S364A+S367A, respectively). For each of the two N protein mutants, two individual molecular clones were isolated and used in this experiment.

The two N protein mutants grow to (slightly or moderately) lower titers in cell but it is not known if (and to what extent) the respective phosphorylations of the nucleocapsid protein affect the cellular transcriptome following infection by HCoV-229E. All experiments were performed at 33°C which is required for efficient replication of this virus.