title: NODULE INCEPTION antagonistically regulates gene expression with nitrate in Lotus japonicus.
abstract: Legumes produce root nodules as symbiotic organs where nitrogen-fixing bacteria are accommodated. Lotus japonicus NODULE INCEPTION (NIN) is an essential factor that specifically and positively regulates nodulation processes, and has evolved from a member of the NIN-like proteins, of which Arabidopsis homologs target nitrate-responsive elements (NREs), and activate gene expression in response to nitrate. It is therefore assumed that the NIN-mediated transcriptional network overlaps with those regulated by NLPs, because of their common DNA-binding RWP-RK domains. However, nodulation is inhibited in the presence of nitrate, and involvement of NIN in nitrate responses has remained largely unknown. Here we determined a consensus of NIN-binding nucleotide sequences (NBSs) by in vitro experiments, and revealed that the sequence pattern was very similar to those of NREs. Chromatin immunoprecitiation (ChIP)-PCR analyses showed that NIN targeted NREs in L. japonicus nitrate-inducible gene promoters, including LjNIR1, LjNRT2.1 and LjNRT2.2. Affinities of NIN binding to the NREs were comparable with that to NBS-yB1a, an NBS on the symbiotic LjNF-YB1 promoter, indicating that NREs are potential targets of NIN. However, rhizobial infection did not activate LjNIR1, LjNRT2.1 and LjNRT2.2. NIN ectopic expression interfered with nitrate-dependent activation of these genes. Nitrate treatment followed by NIN activation down-regulated expression of symbiotic NIN target genes. Our results showed that NIN and nitrate antagonistically regulate expression of genes that are activated by nitrate and NIN, respectively. We propose that this antagonistic relationship prevents inappropriate activation of genes in response to nitrate and rhizobial infection.
authors.affiliation: National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602 Japan. Present address: National Institute for Basic Biology, Okazaki, Aichi, 444-8585 Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Soyano
authors.affiliation: National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602 Japan.
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Shimoda
authors.affiliation: National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602 Japan. Present address: RIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa, 230-0045 Japan. makotoh@affrc.go.jp.
authors.forename: Makoto
authors.initials: M
authors.lastname: Hayashi
mesh_terms: D001483:Base Sequence; D016384:Consensus Sequence / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000187:drug effects*; D017343:Genes, Plant; D000070116:Lotus / Q000187:drug effects / Q000235:genetics* / Q000382:microbiology; D008969:Molecular Sequence Data; D009566:Nitrates / Q000494:pharmacology*; D010940:Plant Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000187:drug effects; D012231:Rhizobium / Q000187:drug effects / Q000502:physiology; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D009566:Nitrates; D010940:Plant Proteins
keywords: Lotus japonicus; NIN-binding nucleotide sequence; NIN-like protein; NODULE INCEPTION; Nitrate; Nitrate-responsive element; Root nodule symbiosis