FRET analysis of in vivo dimerization by RNA-editing enzymes
FRET analysis of in vivo dimerization by RNA-editing enzymes
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DOI：
10.1074/jbc.m511831200
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发表时间：
2006-06-16
影响因子：
4.8
通讯作者：
MacMillan, Andrew M.
中科院分区：
文献类型：
--
作者：
Chilibeck, Kaari A.;Wu, Tao;MacMillan, Andrew M.
ADAR（作用于 RNA 的腺苷脱氨酶）酶家族的成员催化双链 RNA 中腺苷水解脱氨基为肌苷，这是一个人们知之甚少的过程，但对哺乳动物的发育至关重要。我们在转染了带有荧光团的 ADAR1 和 ADAR2 融合蛋白的哺乳动物细胞中进行了荧光共振能量转移实验，以研究这些蛋白之间的关系。这些研究最终证明了 ADAR1 和 ADAR2 的同二聚化，并且还表明 ADAR1 和 ADAR2 在人类细胞中形成异二聚体。表达这些融合蛋白的细胞的 RNase 处理改变了它们的定位，但不影响二聚化。综上所述，这些结果表明同二聚化和异二聚化对于 ADAR 家族成员的体内活性很重要，并且这些关联不依赖于 RNA。
Members of the ADAR (adenosine deaminase that acts on RNA) enzyme family catalyze the hydrolytic deamination of adenosine to inosine within double-stranded RNAs, a poorly understood process that is critical to mammalian development. We have performed fluorescence resonance energy transfer experiments in mammalian cells transfected with fluorophore-bearing ADAR1 and ADAR2 fusion proteins to investigate the relationship between these proteins. These studies conclusively demonstrate the homodimerization of ADAR1 and ADAR2 and also show that ADAR1 and ADAR2 form heterodimers in human cells. RNase treatment of cells expressing these fusion proteins changes their localization but does not affect dimerization. Taken together these results suggest that homo- and heterodimerization are important for the activity of ADAR family members in vivo and that these associations are RNA independent.