Reconstitution of the [4Fe-4S] cluster in FNR and demonstration of the aerobic-anaerobic transcription switch in vitro

被引:155
作者
Green, J [1 ]
Bennett, B [1 ]
Jordan, P [1 ]
Ralph, ET [1 ]
Thomson, AJ [1 ]
Guest, JR [1 ]
机构
[1] UNIV E ANGLIA,SCH CHEM SCI,CTR METALLOPROT SPECT & BIOL,NORWICH NR4 7TJ,NORFOLK,ENGLAND
基金
英国惠康基金;
关键词
D O I
10.1042/bj3160887
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The FNR protein of Escherichia coli is a redox-responsive transcription regulator that activates and represses a family of genes required for anaerobic and aerobic metabolism, Reconstitution of wild-type FNR by anaerobic treatment with ferrous ions, cysteine and the NifS protein of Azotobacter vinelandii leads to the incorporation of two [4Fe-4S](2+) clusters per FNR dimer. The UV-visible spectrum of reconstituted FNR has a broad absorbance at 420 nm, The clusters are EPR silent under anaerobic conditions but are degraded to [3Fe-4S](+) by limited oxidation with air, and completely lost on prolonged air exposure, The association of FNR with the iron-sulphur clusters is confirmed by CD spectroscopy. Incorporation of the [4Fe-4S](2+) clusters increases site-specific DNA binding about 7-fold compared with apo-FNR, Anaerobic transcription activation and repression in vitro likewise depends on the presence of the iron-sulphur cluster: and its inactivation under aerobic conditions provides a demonstration in vitro of the FNR-mediated aerobic-anaerobic transcriptional switch.
引用
收藏
页码:887 / 892
页数:6
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