Py B, Higgins C F, Krisch H M, Carpousis A J
Nuffield Department of Clinical Biochemisstry and Imperial Cancer Research Fund Laboratories, Institute of Molecular Medicine, UK.
Nature. 1996 May 9;381(6578):169-72. doi: 10.1038/381169a0.
The Escherichia coli RNA degradosome is a multi-enzyme complex that contains the exoribonuclease polynucleotide phosphorylase (PNPase) and the endoribonuclease RNase E. Both enzymes are important in RNA processing and messenger RNA degradation. Here we report that enolase and RhlB are two other major components of the degradosome. Enolase is a glycolytic enzyme with an unknown role in RNA metabolism. RhlB is a member of the DEAD-box family of ATP-dependent RNA helicases, which are found in both prokaryotes and eukaryotes. We show that the degradosome has an ATP-dependent activity that aids the degradation of structured RNA by PNPase. Incubation of the degradosome with affinity-purified antibody against RhlB inhibited the ATP-stimulated RNA degradation. These results suggest that RhlB acts by unwinding RNA structures that impede the processive activity of PNPase. RhlB is thus an important enzyme in mRNA turnover.
大肠杆菌RNA降解体是一种多酶复合体,包含外切核糖核酸酶多核苷酸磷酸化酶(PNPase)和内切核糖核酸酶RNase E。这两种酶在RNA加工和信使RNA降解过程中都很重要。在此我们报告烯醇化酶和RhlB是降解体的另外两个主要成分。烯醇化酶是一种糖酵解酶,在RNA代谢中的作用尚不清楚。RhlB是依赖ATP的RNA解旋酶DEAD-box家族的成员,在原核生物和真核生物中均有发现。我们发现降解体具有依赖ATP的活性,可辅助PNPase对结构化RNA的降解。用针对RhlB的亲和纯化抗体孵育降解体可抑制ATP刺激的RNA降解。这些结果表明,RhlB通过解开阻碍PNPase进行性活性的RNA结构来发挥作用。因此,RhlB是mRNA周转过程中的一种重要酶。