One type of RNA editing involves the conversion of adenosine residues into inosine in dsRNA through the action of ADAR (adenosine deaminases acting on RNA). Three ADAR gene family members (ADAR1-3) have been identified in humans and rodents. When it occurs within the coding sequence, A-to-l RNA editing could result in the synthesis of proteins not directly encoded by the genome, as demonstrated with transcripts of GluR ion channels and 5-HT2c serotonin receptors. However, the most common targets for A- to-l editing are non-coding RNAs that contain inverted repeats of repetitive elements located within introns and 3' UTRs. During the past 14 years, this grant has enabled us to clone, for the first time, ADAR1, the first identified member of the ADAR gene family; this in turn led to the identification and cloning of ADAR2 and ADAR3. During the current grant support period, we created an ADAR1 null mutation in mice that causes widespread apoptosis and consequent embryonic lethal phenotypes. Our results suggest that ADAR1 is critically important for survival of numerous tissues by editing a currently unkowntarget dsRNA(s). We also demonstrated that homodimer formation of ADAR is essential for the site-selective A-to-l editing mechanism. Finally, our recent studies revealed that ADAR1 avidly binds siRNA and limits its efficacy, while both ADAR1 and ADAR2 edit specific adenosine residues of certain miRNA precursor dsRNAs, exposing the interaction of the RNAi and RNA editing pathways in mammalian cells. In this renewal application, we propose to: 1) Investigate the functional interaction of the two monomers of homodimeric ADAR for site-selective A-to-l editing and binding of siRNAs using site-directed mutagenesis of the ADAR functional domains, 2) Clone and identify target dsRNAs specifically bound by ADAR1 that may be involved in regulation of apoptosis, 3) Determine critical steps affected by A-to-l RNA editing of miRNA precursors by analyzing in vitro processing of edited miRNA precursors using Drosha and Dicer proteins or their complexes with associated factors and in vivo processing and nuclear export in transfected cells. The overall goal of this project is to elucidate the molecular mechanism of A-to-l RNA editing and to better understand its biological significance. Our research proposal will generate important information on regulation of apoptosis, which is highly relevent to many human diseases including cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM040536-16
Application #
7317366
Study Section
Molecular Genetics A Study Section (MGA)
Program Officer
Tompkins, Laurie
Project Start
1991-07-01
Project End
2009-11-30
Budget Start
2007-12-01
Budget End
2008-11-30
Support Year
16
Fiscal Year
2008
Total Cost
$324,709
Indirect Cost
Name
Wistar Institute
Department
Type
DUNS #
075524595
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Nishikura, Kazuko (2017) Oesophageal cancer: RNA editing of SLC22A3 mRNAs: causative relevance to familial ESCC? Nat Rev Gastroenterol Hepatol 14:569-570
Sakurai, Masayuki; Shiromoto, Yusuke; Ota, Hiromitsu et al. (2017) ADAR1 controls apoptosis of stressed cells by inhibiting Staufen1-mediated mRNA decay. Nat Struct Mol Biol 24:534-543
Tan, Meng How; Li, Qin; Shanmugam, Raghuvaran et al. (2017) Dynamic landscape and regulation of RNA editing in mammals. Nature 550:249-254
Song, Chunzi; Sakurai, Masayuki; Shiromoto, Yusuke et al. (2016) Functions of the RNA Editing Enzyme ADAR1 and Their Relevance to Human Diseases. Genes (Basel) 7:
Miyake, Kotaro; Ohta, Toshio; Nakayama, Hisako et al. (2016) CAPS1 RNA Editing Promotes Dense Core Vesicle Exocytosis. Cell Rep 17:2004-2014
Nishikura, Kazuko (2016) A-to-I editing of coding and non-coding RNAs by ADARs. Nat Rev Mol Cell Biol 17:83-96
Gumireddy, Kiranmai; Li, Anping; Kossenkov, Andrew V et al. (2016) The mRNA-edited form of GABRA3 suppresses GABRA3-mediated Akt activation and breast cancer metastasis. Nat Commun 7:10715
Watanabe, Yoshihisa; Yoshimoto, Kanji; Tatebe, Harutsugu et al. (2014) Enhancement of alcohol drinking in mice depends on alterations in RNA editing of serotonin 2C receptors. Int J Neuropsychopharmacol 17:739-51
Ota, Hiromitsu; Sakurai, Masayuki; Gupta, Ravi et al. (2013) ADAR1 forms a complex with Dicer to promote microRNA processing and RNA-induced gene silencing. Cell 153:575-89
Nishikura, Kazuko; Sakurai, Masayuki; Ariyoshi, Kantaro et al. (2013) Antagonistic and stimulative roles of ADAR1 in RNA silencing. RNA Biol 10:1240-7

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