title: Arabidopsis ANAC069 binds to C[A/G]CG[T/G] sequences to negatively regulate salt and osmotic stress tolerance.
abstract: ANAC069 binds to the DNA sequence of C[A/G]CG[T/G] to regulate the expression of genes, resulting in decreased ROS scavenging capability and proline biosynthesis, which contribute to increased sensitivity to salt and osmotic stress. NAM-ATAF1/2 and CUC2 (NAC) proteins are plant-specific transcription factors that play important roles in abiotic stress responses. In the present study, we characterized the physiological and regulatory roles of Arabidopsis thaliana ANAC069 in response to abiotic stresses. Arabidopsis plants overexpressing ANAC069 displayed increased sensitivity to abscisic acid, salt, and osmotic stress. Conversely, ANAC069 knockdown plants showed enhanced tolerance to salt and osmotic stress, but no change in ABA sensitivity. Further studies showed that ANAC069 inhibits the expression of SOD, POD, GST, and P5CS genes. Consequently, the transcript level of ANAC069 correlated negatively with the reactive oxygen species (ROS) scavenging ability and the proline level. The genes regulated by ANAC069 were further studied using a gene chip on a genome-wide scale, and 339 and 226 genes up- and downregulated by ANAC069 were identified. Analysis of the promoters of the genes affected by ANAC069 suggested that ANAC069 regulates the expression of genes mainly through interacting with the DNA sequence C[A/G]CG[T/G] in response to abiotic stresses. Collectively, our data suggest that ANAC069 could recognize C[A/G]CG[T/G] sequences to regulate the expression of genes that negatively regulates salt and osmotic stress tolerance by decreasing ROS scavenging capability and proline biosynthesis.
authors.affiliation: Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, 830011, Urumqi, China.|Agricultural College, Heilongjiang Bayi Agricultural University, 163319, Daqing, China.
authors.forename: Lin
authors.initials: L
authors.lastname: He
authors.affiliation: Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, 830011, Urumqi, China.|State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: Xinxin
authors.initials: X
authors.lastname: Shi
authors.affiliation: Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, 830011, Urumqi, China.|State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: Yanmin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, 830011, Urumqi, China.|State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Agricultural College, Heilongjiang Bayi Agricultural University, 163319, Daqing, China.
authors.forename: Kejun
authors.initials: K
authors.lastname: Yang
authors.affiliation: Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, 830011, Urumqi, China. wangyucheng@ms.xjb.ac.cn.|State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China. wangyucheng@ms.xjb.ac.cn.
authors.forename: Yucheng
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000040:Abscisic Acid / Q000494:pharmacology; D000222:Adaptation, Physiological / Q000187:drug effects / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D009997:Osmotic Pressure; D010937:Plant Growth Regulators / Q000494:pharmacology; D030821:Plants, Genetically Modified; D011392:Proline / Q000096:biosynthesis; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055049:Salt Tolerance / Q000187:drug effects / Q000235:genetics*; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D014867:Water / Q000378:metabolism
chemical_list: C000602371:ANAC069 protein, Arabidopsis; D029681:Arabidopsis Proteins; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D010937:Plant Growth Regulators; D017382:Reactive Oxygen Species; D014867:Water; D012965:Sodium Chloride; D000040:Abscisic Acid; D011392:Proline; D013482:Superoxide Dismutase
keywords: ANAC069; Abiotic stress; Cis-acting element; Gene expression; Proline biosynthesis; Reactive oxygen species