Gaymard F, Boucherez J, Briat J F
Centre National de la Recherche Scientifique (Unité de Recherche 2133), Institut National de la Recherche Agronomique, Montpellier, France.
Biochem J. 1996 Aug 15;318 ( Pt 1)(Pt 1):67-73. doi: 10.1042/bj3180067.
A ferritin cDNA, AtFer1, from seedlings of Arabidopsis thaliana has been characterized. The deduced amino acid sequence of the AtFer1 protein indicates that A. thaliana ferritin shares the same characteristics as the plant ferritin already characterized from the Leguminosae and Graminacea families: (i) it contains an additional sequence in its N-terminal part composed of two domains: a transit peptide responsible for plastid targeting and an extension peptide; (ii) amino acids that form the ferroxidase centre of H-type animal ferritin, as well as Glu residues characteristic of L-type animal ferritin, are conserved in AtFer1; (iii) the C-terminal part of the A. thaliana ferritin subunit defining the E-helix is divergent from its animal counterpart, and confirms that 4-fold-symmetry axis channels are hydrophilic in plant ferritin. Southern blot experiments indicate that AtFer1 is likely to be encoded by a unique gene in the A. thaliana genome, although a search in the NCBI dbEST database indicates that other ferritin genes, divergent from AtFer1, may exist. Iron loading of A. thaliana plantlets increased ferritin mRNA and protein abundance. In contrast to maize, the transcript abundance of a gene responding to abscisic acid (RAB18) did not increase in response to iron loading treatment, and A. thaliana ferritin mRNA abundance is not accumulated in response to a treatment with exogenous abscisic acid, at least in the culture system used in this study. In addition, iron-induced increases in ferritin mRNA abundance were the same as wild-type plants in abi1 and abi2 mutants of A. thaliana, both affected in the abscisic acid response in vegetative tissues. Increased AtFer1 transcript abundance in response to iron is inhibited by the antioxidant N-acetylcysteine. These results indicate that an oxidative pathway, independent of abscisic acid, could be responsible for the iron induction of ferritin synthesis in A. thaliana.
已对来自拟南芥幼苗的铁蛋白cDNA(AtFer1)进行了表征。AtFer1蛋白的推导氨基酸序列表明,拟南芥铁蛋白与豆科和禾本科已表征的植物铁蛋白具有相同的特征:(i)其N端部分包含一个由两个结构域组成的额外序列:负责质体靶向的转运肽和延伸肽;(ii)形成H型动物铁蛋白铁氧化酶中心的氨基酸以及L型动物铁蛋白特有的Glu残基在AtFer1中保守;(iii)拟南芥铁蛋白亚基定义E螺旋的C端部分与其动物对应物不同,并证实4重对称轴通道在植物铁蛋白中是亲水的。Southern印迹实验表明,AtFer1可能由拟南芥基因组中的一个独特基因编码,尽管在NCBI dbEST数据库中的搜索表明可能存在与AtFer1不同的其他铁蛋白基因。拟南芥幼苗的铁加载增加了铁蛋白mRNA和蛋白质丰度。与玉米不同,响应脱落酸(RAB18)的基因转录本丰度在铁加载处理后没有增加,并且拟南芥铁蛋白mRNA丰度在至少本研究使用的培养系统中对外源脱落酸处理没有积累。此外,在拟南芥的abi1和abi2突变体中,铁诱导的铁蛋白mRNA丰度增加与野生型植物相同,这两个突变体在营养组织中的脱落酸反应均受到影响。抗氧化剂N-乙酰半胱氨酸抑制了铁诱导的AtFer1转录本丰度增加。这些结果表明,一条独立于脱落酸的氧化途径可能负责拟南芥中铁诱导的铁蛋白合成。