Aerobic-type ribonucleotide reductase in the anaerobe Bacteroides fragilis.
Aerobic-type ribonucleotide reductase in the anaerobe Bacteroides fragilis.
复制标题
厌氧脆弱拟杆菌中的需氧型核糖核苷酸还原酶。
DOI：
10.1128/jb.184.4.895-903.2002
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发表时间：
2002
影响因子：
3.2
通讯作者：
Smith,CJeffrey
中科院分区：
文献类型：
--
作者：
Smalley,Darren;Rocha,EdsonR;Smith,CJeffrey
Bacteroides fragilis, a component of the normal intestinal flora, is an obligate anaerobe capable of long-term survival in the presence of air. Survival is attributed to an elaborate oxidative stress response that controls the induction of more than 28 peptides, but there is limited knowledge concerning the identities of these peptides. In this report, RNA fingerprinting by arbitrarily primed PCR identified five new genes whose expression increased following exposure to O2. Nucleotide sequence analysis of the cloned genes indicated that they encoded an outer membrane protein, an aspartate decarboxylase, an efflux pump, heat shock protein HtpG, and an NrdA ortholog constituting the large subunit of a class Ia ribonucleotide reductase (RRase). Attention was focused on thenrdAgene since class I RRases are obligate aerobic enzymes catalyzing the reduction of ribonucleoside 5′-diphosphates by a mechanism that requires molecular oxygen for activity. Sequence analysis of thenrdlocus showed that two genes,nrdAandnrdB, are located in the same orientation in a 4.5-kb region. Northern hybridization and primer extension experiments confirmed induction of the genes by O2and suggested they are an operon. TheB. fragilis nrdAandnrdBgenes were overexpressed inEscherichia coli, and CDP reductase assays confirmed that they encoded an active enzyme. The enzyme activity was inhibited by hydroxyurea, and ATP was shown to be a positive effector of CDP reductase activity, while dATP was an inhibitor, indicating that the enzyme was a class Ia RRase. AnrdAmutant was viable under anaerobic conditions but had decreased survival following exposure to O2, and it could not rapidly resume growth after O2treatment. The results presented indicate that during aerobic conditionsB. fragilisNrdAB may have a role in maintaining deoxyribonucleotide pools for DNA repair and growth recovery.
