SARS-CoV-2 open reading frame 3c protein found to be targeted selectively to mitochondria biorxivpreprint Cambridge_Uni helsinkiuni SARSCoV2 COVID19 Mitochondria
By Pooja Toshniwal PahariaNov 17 2022Reviewed by Aimee Molineux In a recent study posted to the bioRxiv* preprint server, researchers presented a functional analysis of the severe acute respiratory syndrome coronavirus 2 open reading frame 3c protein.
For testing the hypothesis and measuring the protein expression levels, an expression cassette extending from the 5′ end of ORF3a sgmRNA transcript until the final coding nucleotide of the protein was created. GenBank sequence records of ORF3c sequences were inspected. Multiple mutants were created that ablated the protein's initial and subsequent AUG codons, ORF3a AUG codons, or ORF3c AUG.
To assess the membrane topology of ORF3c, the protein was modified to enable identification of ER import, and an OPG2 tag was incorporated at the C- or N-terminal ends. Radiolabelled ORF3c was synthesized, and the endoplasmic reticulum membranes of OPG2-tagged ORF3c variants were subjected to endoglycosidase H treatment to detect N-glycosylation.
Results Genetics & Genomics eBook Compilation of the top interviews, articles, and news in the last year. Download a copy today ORF3c, a conserved and small protein, overlapped the ORF3a protein in the +1 reading frame and coincided precisely with an area of significantly high synonymous site conservation in ORF3a-frame codons, indicating that the ORF3c gene was a functional SARS-CoV-2 gene.
Therefore, nearly half of the pre-initiation complexes scanned past the first and second AUGs of ORF3a sgmRNA and initiated at the next AUG downstream for ORF3c translation. ORF3c did not multimerize and was not likely to form ionic channels.
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