The Long-Term Evolutionary History of Gradual Reduction of CpG Dinucleotides in the SARS-CoV-2 Lineage

SIMPLE SUMMARY: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the coronavirus disease 2019 (COVID-19), a pandemic that infected over 81 million people worldwide. This has led the scientific community to characterize the genome of this virus, including its nucleotide composition. Investigation of the dinucleotide frequency revealed that the proportion of CG dinucleotides (CpG) is highly reduced in the viral genomes. Since CpG dinucleotides is the target site for the host antiviral zinc finger protein, it has been suggested that the reduction in the proportion of CpG is the viral response to escape from the host defense machinery. In the present study, we investigated the time of origin of reduction in the CpG content. Whole genome analyses based on all representative viral genomes of the group Betacoronavirus revealed that the CpG content in the lineage of SARS-CoV-2 has been progressively declining over the past 1213 years. The depletion of CpG was found to occur at neutral—as well as selectively constrained—positions of the viral genomes. ABSTRACT: Recent studies suggested that the fraction of CG dinucleotides (CpG) is severely reduced in the genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The CpG deficiency was predicted to be the adaptive response of the virus to evade degradation of the viral RNA by the antiviral zinc finger protein that specifically binds to CpG nucleotides. By comparing all representative genomes belonging to the genus Betacoronavirus, this study examined the potential time of origin of CpG depletion. The results of this investigation revealed a highly significant correlation between the proportions of CpG nucleotide (CpG content) of the betacoronavirus species and their times of divergence from SARS-CoV-2. Species that are distantly related to SARS-CoV-2 had much higher CpG contents than that of SARS-CoV-2. Conversely, closely related species had low CpG contents that are similar to or slightly higher than that of SARS-CoV-2. These results suggest a systematic and continuous reduction in the CpG content in the SARS-CoV-2 lineage that might have started since the Sarbecovirus + Hibecovirus clade separated from Nobecovirus, which was estimated to be 1213 years ago. This depletion was not found to be mediated by the GC contents of the genomes. Our results also showed that the depletion of CpG occurred at neutral positions of the genome as well as those under selection. The latter is evident from the progressive reduction in the proportion of arginine amino acid (coded by CpG dinucleotides) in the SARS-CoV-2 lineage over time. The results of this study suggest that shedding CpG nucleotides from their genome is a continuing process in this viral lineage, potentially to escape from their host defense mechanisms.