Shown above are sequence traces from the same region of genomic DNA and cDNAs fr

Question

Shown above are sequence traces from the same region of genomic DNA and cDNAs from cell lines that were untreated (siRNA control) or treated with siADAR1 or siADAR2. (I won't lie; these editing sites are actually in the 3'UTR of this pre-mRNA. But for now, we wall pretend that they are in the coding region of the gene.) The left sequencing trace shows the sequence of genomic DNA. The next trace (si control) shows the same region from cDNA from a wild type (normal) cell. The third and fourth show traces from the same region of the cDNAs from ADAR1 and ARAD2 knockdown cells, respectively. a. The traces show as G for the control. The nucleotide that is actually present in the mRNA is _____ b. If we don't know the position in the codon, the A that is sometimes read as G could be in the first, second or third position, or CAA, AAG or AGA. What are the possible amino acid changes that would result for each of these possibilities? Please show how you arrived at your answer. c. Does the observed RNA editing require ADAR1, ADAR2, neither, or both? Please explain your answer briefly.

Explanation / Answer

a) The nucleotide to be present in the mRNA is C- cytosine, Since G triple bonds with C according to complementary base pairing rule.

b) The amino acids could be Gln - Lys - Arg for respective codon, based on codon table.

c) Editing is required in siADAR1 cDNA, since there is a nucleotide substitution of A instead of G in comparison with the control.

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