Ehrt S, Ornston L N, Hillen W
Lehrstuhl für Mikrobiologie, Friedrich-Alexander Universität Erlangen-Nürnberg, Germany.
J Bacteriol. 1994 Jun;176(12):3493-9. doi: 10.1128/jb.176.12.3493-3499.1994.
Members of the sigma 54 protein family, encoded by rpoN, are required for the transcription of genes associated with specialized metabolic functions. The ability to grow with phenol appears to be a specialized trait because it is expressed by few of the microorganisms that grow with catechol, the metabolic product of phenol monooxygenase. A mutation preventing the expression of phenol monooxygenase in the bacterial strain Acinetobacter calcoaceticus NCIB8250 was complemented by wild-type DNA segments containing an open reading frame encoding a member of the sigma 54 protein family. DNA sequencing revealed a second open reading frame, designated ORF2, directly downstream of A. calcoaceticus rpoN. The locations of both ORF2 and the 113-residue amino acid sequence of its product are highly conserved in other bacteria. The mutation preventing the expression of rpoN results in an opal codon that terminates the translation of RpoN at a position corresponding to Trp-91 in the 483-residue amino acid sequence of the wild-type protein. Negative autoregulation of rpoN was suggested by the fact that the mutation inactivating RpoN enhanced the transcription of rpoN. Primer extension revealed independent transcription start sites for rpoN and ORF2.
由rpoN编码的σ54蛋白家族成员是与特殊代谢功能相关基因转录所必需的。利用苯酚生长的能力似乎是一种特殊性状,因为在以苯酚单加氧酶的代谢产物儿茶酚生长的微生物中,只有少数能够表达这种能力。在醋酸钙不动杆菌NCIB8250菌株中,一个阻止苯酚单加氧酶表达的突变被含有编码σ54蛋白家族成员的开放阅读框的野生型DNA片段所互补。DNA测序揭示了一个直接位于醋酸钙不动杆菌rpoN下游的第二个开放阅读框,命名为ORF2。ORF2及其产物的113个氨基酸序列在其他细菌中高度保守。阻止rpoN表达的突变产生了一个乳白密码子,该密码子在与野生型蛋白483个氨基酸序列中第91位色氨酸相对应的位置终止RpoN的翻译。失活RpoN的突变增强了rpoN的转录,这一事实表明rpoN存在负自调控。引物延伸揭示了rpoN和ORF2独立的转录起始位点。