RNA editing of the Q/R site of GluA2 in different cultured cell lines that constitutively express different levels of RNA editing enzyme ADAR2

Adenosine deaminase acting on RNA 2 (ADAR2) catalyzes RNA editing at the glutamine/arginine (Q/R) site of GluA2, and an ADAR2 deficiency may play a role in the death of motor neurons in ALS patients. The expression level of ADAR2 mRNA is a determinant of the editing activity at the GluA2 Q/R site in human brain but not in cultured cells. Therefore, we investigated the extent of Q/R site-editing in the GluA2 mRNA and pre-mRNA as well as the ADAR2 mRNA and GluA2 mRNA and pre-mRNA levels in various cultured cell lines. The extent of the GluA2 mRNA editing was 100% except in SH-SY5Y cells, which have a much lower level of ADAR2 than the other cell lines examined. The ADAR2 activity at the GluA2 pre-mRNA Q/R site correlated with the ADAR2 mRNA level relative to the GluA2 pre-mRNA. SH-SY5Y cells expressed higher level of the GluA2 mRNA in the cytoplasm compared with other cell lines. These results suggest that the ADAR2 expression level reflects editing activity at the GluA2 Q/R site and that although the edited GluA2 pre-mRNA is readily spliced, the unedited GluA2 pre-mRNA is also spliced and transported to the cytoplasm when ADAR2 expression is low.

► ADAR2 catalyzes RNA editing at the Q/R site in the GluA2 pre-mRNA.
► The edited GluA2 pre-mRNA was more efficiently spliced than the unedited pre-mRNA.
► The GluA2 mRNA was not always edited at the Q/R site in cultured cell lines.
► ADAR2 activity correlated with the abundance of ADAR2 mRNA relative to GluA2 pre-mRNA.
► Both edited and unedited GluA2 mRNAs were equally transported to the cytoplasm.