An ADAR that edits transcripts encoding ion channel subunits functions as a dimer

被引:94
作者
Gallo, A [1 ]
Keegan, LP [1 ]
Ring, GM [1 ]
O'Connell, MA [1 ]
机构
[1] Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
关键词
ADAR; dimer; Drosophila; ion channels; RNA editing;
D O I
10.1093/emboj/cdg327
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In this report, we establish that Drosophila ADAR (adenosine deaminase acting on RNA) forms a dimer on double-stranded (ds) RNA, a process essential for editing activity. The minimum region required for dimerization is the N-terminus and dsRNA-binding domain 1 (dsRBD1). Single point mutations within dsRBD1 abolish RNA-binding activity and dimer formation. These mutations and glycerol gradient analysis indicate that binding to dsRNA is important for dimerization. However, dimerization can be uncoupled from dsRNA-binding activity, as a deletion of the N-terminus (amino acids 1-46) yields a monomeric ADAR that retains the ability to bind dsRNA but is inactive in an editing assay, demonstrating that ADAR is only active as a dimer. Different isoforms of ADAR with different editing activities can form heterodimers and this can have a significant effect on editing in vitro as well as in vivo. We propose a model for ADAR dimerization whereby ADAR monomers first contact dsRNA; however, it is only when the second monomer binds and a dimer is formed that deamination occurs.
引用
收藏
页码:3421 / 3430
页数:10
相关论文
共 54 条
[1]  
[Anonymous], 1989, MOL CLONING LAB MANU
[2]   RNA-editing of the 5-HT2C receptor alters agonist-receptor-effector coupling specificity [J].
Berg, KA ;
Cropper, JD ;
Niswender, CM ;
Sanders-Bush, E ;
Emeson, RB ;
Clarke, WP .
BRITISH JOURNAL OF PHARMACOLOGY, 2001, 134 (02) :386-392
[3]   CYTIDINE DEAMINASE - THE 2-CENTER-DOT-3-ANGSTROM CRYSTAL-STRUCTURE OF AN ENZYME - TRANSITION-STATE ANALOG COMPLEX [J].
BETTS, L ;
XIANG, SB ;
SHORT, SA ;
WOLFENDEN, R ;
CARTER, CW .
JOURNAL OF MOLECULAR BIOLOGY, 1994, 235 (02) :635-656
[4]   Regulation of serotonin-2C receptor G-protein coupling by RNA editing [J].
Burns, CM ;
Chu, H ;
Rueter, SM ;
Hutchinson, LK ;
Canton, H ;
SandersBush, E ;
Emeson, RB .
NATURE, 1997, 387 (6630) :303-308
[5]   NMR SOLUTION STRUCTURE OF A DSRNA BINDING DOMAIN FROM DROSOPHILA STAUFEN PROTEIN REVEALS HOMOLOGY TO THE N-TERMINAL DOMAIN OF RIBOSOMAL-PROTEIN S5 [J].
BYCROFT, M ;
GRUNERT, S ;
MURZIN, AG ;
PROCTOR, M ;
STJOHNSTON, D .
EMBO JOURNAL, 1995, 14 (14) :3563-3571
[6]   Alternative splicing:: multiple control mechanisms and involvement in human disease [J].
Cáceres, JF ;
Kornblihtt, AR .
TRENDS IN GENETICS, 2002, 18 (04) :186-193
[7]   Potential Alu function: Regulation of the activity of double-stranded RNA-activated kinase PKR [J].
Chu, WM ;
Ballard, R ;
Carpick, BW ;
Williams, BRG ;
Schmid, CW .
MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (01) :58-68
[8]   DOUBLE-STRANDED-RNA-DEPENDENT PROTEIN-KINASE AND TAR RNA-BINDING PROTEIN FORM HOMODIMERS AND HETERODIMERS IN-VIVO [J].
COSENTINO, GP ;
VENKATESAN, S ;
SERLUCA, FC ;
GREEN, SR ;
MATHEWS, MB ;
SONENBERG, N .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (21) :9445-9449
[9]   Proteins binding to duplexed RNA: one motif, multiple functions [J].
Fierro-Monti, I ;
Mathews, MB .
TRENDS IN BIOCHEMICAL SCIENCES, 2000, 25 (05) :241-246
[10]  
Gerber A, 1997, RNA, V3, P453