Laudenbach D E, Straus N A
Department of Botany, University of Toronto, Ontario, Canada.
J Bacteriol. 1988 Nov;170(11):5018-26. doi: 10.1128/jb.170.11.5018-5026.1988.
Recently we have reported that the flavodoxin gene from the cyanobacterium Anacystis nidulans R2 is transcribed as part of an iron stress-induced operon containing multiple mRNA species (D. E. Laudenbach, M. E. Reith, and N. A. Straus, J. Bacteriol. 170: 258-265, 1988). Here we report that nucleotide sequence analyses of DNA located immediately upstream of the flavodoxin gene revealed an open reading frame of 1,026 bases (designated isiA; iron stress inducible) with a deduced amino acid sequence showing similarity to that of the psbC polypeptide of higher plants and cyanobacteria. Assuming proteolytic cleavage of the initial methionine residue, the open reading frame encodes a 341-amino-acid polypeptide with a molecular mass of 36,824 daltons. Amino acid sequence comparisons with known psbC polypeptides from spinach and A. nidulans R2 showed extensive similarity, especially in the proposed membrane-spanning regions. Mung bean nuclease mapping and primer extension experiments have localized a transcriptional start site to a position 19 bases upstream from the first methionine codon of the isiA gene product. The upstream region contains an Escherichia coli-like -10 sequence but lacks the typical -35 consensus sequence. Approximately 15, 25, and 150 bases upstream from the isiA transcription start site are 17 base sequences which resemble the operator sequences of iron-regulated genes of E. coli.
最近我们报道,来自蓝细菌集胞藻6803 R2的黄素氧还蛋白基因作为一个铁应激诱导操纵子的一部分被转录,该操纵子包含多个mRNA种类(D. E. 劳登巴赫、M. E. 赖思和N. A. 施特劳斯,《细菌学杂志》170: 258 - 265, 1988)。在此我们报道,对黄素氧还蛋白基因紧邻上游的DNA进行的核苷酸序列分析揭示了一个1026个碱基的开放阅读框(命名为isiA;铁应激诱导),其推导的氨基酸序列与高等植物和蓝细菌的psbC多肽的氨基酸序列相似。假设起始甲硫氨酸残基被蛋白酶切割,该开放阅读框编码一个341个氨基酸的多肽,分子量为36,824道尔顿。与菠菜和集胞藻6803 R2已知的psbC多肽进行的氨基酸序列比较显示出广泛的相似性,尤其是在推测的跨膜区域。绿豆核酸酶图谱分析和引物延伸实验已将转录起始位点定位到isiA基因产物第一个甲硫氨酸密码子上游19个碱基的位置。上游区域含有一个类似大肠杆菌的-10序列,但缺乏典型的-35共有序列。在isiA转录起始位点上游约15、25和150个碱基处是17个碱基的序列,它们类似于大肠杆菌铁调节基因的操纵序列。