title: Posttranslational regulation of the iron deficiency basic helix-loop-helix transcription factor FIT is affected by iron and nitric oxide.
abstract: Understanding iron (Fe) sensing and regulation is important for targeting key genes for important nutritional traits like Fe content. The basic helix-loop-helix transcription factor FIT (for FER-LIKE FE DEFICIENCY-INDUCED TRANSCRIPTION FACTOR) controls Fe acquisition genes in dicot roots. Posttranscriptional regulation of transcription factors allows rapid adaptation to cellular changes and was also described for FIT. However, the mechanisms behind this regulation of FIT were for a long time not known. Here, we studied the posttranscriptional control mechanisms of FIT in Arabidopsis (Arabidopsis thaliana) and identified nitric oxide as a stabilizing stimulus for FIT protein abundance. Using cycloheximide, we confirmed that the level of FIT protein was regulated by way of protein turnover in wild-type and hemagglutinin-FIT plants. Upon cycloheximide treatment, FIT activity was hardly compromised, since Fe deficiency genes like IRON-REGULATED TRANSPORTER1 and FERRIC REDUCTASE OXIDASE2 were still inducible by Fe deficiency. A small pool of "active" FIT was sufficient for the induction of Fe deficiency downstream responses. Nitric oxide inhibitors caused a decrease of FIT protein abundance and, in the wild type, also a decrease in FIT activity. This decrease of FIT protein levels was reversed by the proteasomal inhibitor MG132, suggesting that in the presence of nitric oxide FIT protein was less likely to be a target of proteasomal degradation. Independent of FIT transcription, FIT protein stability and FIT protein activity, therefore, were targets of control mechanisms in response to Fe and nitric oxide. We summarize our results in a model that explains the different steps of FIT regulation integrating the plant signals that control FIT.
authors.affiliation: Department of Biosciences-Plant Biology, Saarland University, D-66123 Saarbrucken, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Meiser
authors.forename: Sivasenkar
authors.initials: S
authors.lastname: Lingam
authors.forename: Petra
authors.initials: P
authors.lastname: Bauer
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D003513:Cycloheximide / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant / Q000187:drug effects*; D007501:Iron / Q000378:metabolism / Q000494:pharmacology*; D007976:Leupeptins / Q000494:pharmacology; D051379:Mice; D008954:Models, Biological; D009569:Nitric Oxide / Q000494:pharmacology*; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D036226:Seedlings / Q000187:drug effects / Q000235:genetics / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D051792:Basic Helix-Loop-Helix Proteins; D003513:Cycloheximide; D015853:Cysteine Proteinase Inhibitors; D007501:Iron; D007976:Leupeptins; D009569:Nitric Oxide; D046988:Proteasome Endopeptidase Complex; D011500:Protein Synthesis Inhibitors; C501411:FIT1 protein, Arabidopsis; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde