Requirement of dimerization for RNA editing activity of adenosine deaminases acting on RNA.
Adenosine deaminases acting on RNA (ADAR) convert adenosine residues into inosines in double-stranded RNA. Three vertebrate ADAR gene family members, ADAR1, ADAR2, and ADAR3, have been identified. The catalytic domain of all three ADAR gene family members is very similar to that of Escherichia coli cytidine deaminase and APOBEC-1. Homodimerization is essential for the enzyme activity of those cytidine deaminases. In this study, we investigated the formation of complexes between differentially epitope-tagged ADAR monomers by sequential affinity chromatography and size exclusion column chromatography. Both ADAR1 and ADAR2 form a stable enzymatically active homodimer complex, whereas ADAR3 remains as a monomeric, enzymatically inactive form. No heterodimer complex formation among different ADAR gene family members was detected. Analysis of HeLa and mouse brain nuclear extracts suggested that endogenous ADAR1 and ADAR2 both form a homodimer complex. Interestingly, endogenous ADAR3 also appears to form a homodimer complex, indicating the presence of a brain-specific mechanism for ADAR3 dimerization. Homodimer formation may be necessary for ADAR to act as active deaminases. Analysis of dimer complexes consisting of one wild-type and one mutant monomer suggests functional interactions between the two subunits during site-selective RNA editing.
Cho
Dan-Sung C
DS
Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
Yang
Weidong
W
Lee
Joshua T
JT
Shiekhattar
Ramin
R
Murray
John M
JM
Nishikura
Kazuko
K
D000243:Adenosine Deaminase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D019281:Dimerization; D004926:Escherichia coli; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D012313:RNA / Q000235:genetics / Q000378:metabolism; D017393:RNA Editing; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D012313:RNA; D000243:Adenosine Deaminase