Knockout of GMC-oxidoreductase genes reveals functional redundancy in mimivirus

Alempic, J.-M.; Bisio, H.; Villalta, A.; Santini, S.; Lartigue, A.; Schmitt, A.; Bugnot, C.; Notaro, A.; BELMUDES, L.; Adrait, A.; Poirot, O.; Ptchelkine, D.; De Castro, C.; Coute, Y.; Abergel, C.

doi:10.1101/2023.04.28.538727

bioRxiv · 10.1101/2023.04.28.538727

Knockout of GMC-oxidoreductase genes reveals functional redundancy in mimivirus

Alempic, J.-M.·Bisio, H.·Villalta, A.·Santini, S.·Lartigue, A.·Schmitt, A.·Bugnot, C.·Notaro, A.·BELMUDES, L.·Adrait, A.·Poirot, O.·Ptchelkine, D.·De Castro, C.·Coute, Y.·Abergel, C.

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Abstract

The mimivirus 1.2Mb genome was shown to be organized into a nucleocapsid-like genomic fiber encased in the nucleoid compartment inside the icosahedral capsid (1). The genomic fiber protein shell is composed of a mixture of two GMC-oxidoreductase paralogs, one of them being the main component of the glycosylated layer of fibrils at the surface of the virion (2). In this study, we determined the effect of the deletion of each of the corresponding genes on the genomic fiber and the layer of surface fibrils. First, we deleted the GMC-oxidoreductase the most abundant in the genomic fiber, and determined its structure and composition in the mutant. As expected, it was composed of the second GMC-oxidoreductase and contained 5- and 6-start helices similar to the wild-type fiber. This result led us to propose a model explaining their coexistence. Then, we deleted the GMC-oxidoreductase the most abundant in the layer of fibrils to analyze its protein composition in the mutant. Second, we showed that the fitness of single mutants and the double mutant were not decreased compared to the wild-type viruses in laboratory conditions. Third, we determined that deleting the GMC-oxidoreductase genes did not impact the glycosylation or the glycan composition of the layer of surface fibrils, despite modifying their protein composition. Since the glycosylation machinery and glycan composition of members of different clades are different (3, 4), we expanded the analysis of the protein composition of the layer of fibrils to members of the B and C clades and showed that it was different among the three clades and even among isolates within the same clade. Taken together, the results obtained on two distinct central processes (genome packaging and virion coating) illustrate an unexpected functional redundancy in members of the family Mimiviridae, suggesting this may be the major evolutionary force behind their giant genomes. One-Sentence SummaryFunctional redundancy preserves mimivirus genomic fiber and layer of fibrils formation.

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Alempic, J.-M., Bisio, H., Villalta, A., Santini, S., Lartigue, A., Schmitt, A., Bugnot, C., Notaro, A., BELMUDES, L., Adrait, A., Poirot, O., Ptchelkine, D., De Castro, C., Coute, Y., Abergel, C.. 2023-04-28. Knockout of GMC-oxidoreductase genes reveals functional redundancy in mimivirus. https://doi.org/10.1101/2023.04.28.538727

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