Mutations in the NB-ARC domain of I-2 that impair ATP hydrolysis cause autoactivation

被引:228
作者
Tameling, WIL
Vossen, JH
Albrecht, M
Lengauer, T
Berden, JA
Haring, MA
Cornelissen, BJC
Takken, FLW
机构
[1] Univ Amsterdam, Swammerdam Inst Life Sci, NL-1090 GB Amsterdam, Netherlands
[2] Max Planck Inst Informat, D-66123 Saarbrucken, Germany
关键词
D O I
10.1104/pp.105.073510
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Resistance (R) proteins in plants confer specificity to the innate immune system. Most R proteins have a centrally located NB-ARC (nucleotide-binding adaptor shared by APAF-1, R proteins, and CED-4) domain. For two tomato (Lycopersicon esculentum) R proteins, I-2 and Mi-1, we have previously shown that this domain acts as an ATPase module that can hydrolyze ATP in vitro. To investigate the role of nucleotide binding and hydrolysis for the function of I-2 in planta, specific mutations were introduced in conserved motifs of the NB-ARC domain. Two mutations resulted in autoactivating proteins that induce a pathogen-independent hypersensitive response upon expression in planta. These mutant forms of I-2 were found to be impaired in ATP hydrolysis, but not in ATP binding, suggesting that the ATP-rather than the ADP-bound state of I-2 is the active form that triggers defense signaling. In addition, upon ADP binding, the protein displayed an increased affinity for ADP suggestive of a change of conformation. Based on these data, we propose that the NB-ARC domain of I-2, and likely of related R proteins, functions as a molecular switch whose state (on/off) depends on the nucleotide bound (ATP/ADP).
引用
收藏
页码:1233 / 1245
页数:13
相关论文
共 77 条
[1]   Disease-associated variants in PYPAF1 and NOD2 result in similar alterations of conserved sequence [J].
Albrecht, M ;
Lengauer, T ;
Schreiber, S .
BIOINFORMATICS, 2003, 19 (17) :2171-2175
[2]   Structural localization of disease-associated sequence variations in the NACHT and LRR domains of PYPAF1 and NOD2 [J].
Albrecht, M ;
Domingues, FS ;
Schreiber, S ;
Lengauer, T .
FEBS LETTERS, 2003, 554 (03) :520-528
[3]   Update on the domain architectures of NLRs and R proteins [J].
Albrecht, M ;
Takken, FLW .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2006, 339 (02) :459-462
[4]   Inactivation of the flax rust resistance gene M associated with loss of a repeated unit within the leucine-rich repeat coding region [J].
Anderson, PA ;
Lawrence, GJ ;
Morrish, BC ;
Ayliffe, MA ;
Finnegan, EJ ;
Ellis, JG .
PLANT CELL, 1997, 9 (04) :641-651
[5]   SCOP database in 2004: refinements integrate structure and sequence family data [J].
Andreeva, A ;
Howorth, D ;
Brenner, SE ;
Hubbard, TJP ;
Chothia, C ;
Murzin, AG .
NUCLEIC ACIDS RESEARCH, 2004, 32 :D226-D229
[6]  
Apweiler R, 2004, NUCLEIC ACIDS RES, V32, pD115, DOI [10.1093/nar/gkw1099, 10.1093/nar/gkh131]
[7]   A novel family of P-loop NTPases with an unusual phyletic distribution and transmembrane segments inserted within the NTPase domain [J].
Aravind, L ;
Iyer, LM ;
Leipe, DD ;
Koonin, EV .
GENOME BIOLOGY, 2004, 5 (05)
[8]   Analysis of alternative transcripts of the flax L6 rust resistance gene [J].
Ayliffe, MA ;
Frost, DV ;
Finnegan, EJ ;
Lawrence, GJ ;
Anderson, PA ;
Ellis, JG .
PLANT JOURNAL, 1999, 17 (03) :287-292
[9]  
Bateman A, 2004, NUCLEIC ACIDS RES, V32, pD138, DOI [10.1093/nar/gkp985, 10.1093/nar/gkr1065, 10.1093/nar/gkh121]
[10]   Plant disease resistance protein signaling: NBS-LRR proteins and their partners [J].
Belkhadir, Y ;
Subramaniam, R ;
Dangl, JL .
CURRENT OPINION IN PLANT BIOLOGY, 2004, 7 (04) :391-399