ribonucleotide reductases of salmonella typhimurium transcriptional regulation and differential role in pathogenesis鼠伤寒沙门氏菌的核苷酸还原酶转录调控和微分在发病机制中的作用.pdfVIP

Ribonucleotide Reductases of Salmonella Typhimurium: Transcriptional Regulation and Differential Role in Pathogenesis ¨ ¤a ¤b Anaıs Panosa , Ignasi Roca* , Isidre Gibert* ` ` ` Institut de Biotecnologia i de Biomedicina and Departament de Genetica i de Microbiologia, Universitat Autonoma de Barcelona, Bellaterra (Cerdanyola del Valles), Barcelona, Spain Abstract Ribonucleotide reductases (RNRs) are essential enzymes that carry out the de novo synthesis of deoxyribonucleotides by reducing ribonucleotides. There are three different classes of RNRs (I, II and III), all having different oxygen dependency and biochemical characteristics. Salmonella enterica serovar Typhimurium (S. Typhimurium) harbors class Ia, class Ib and class III RNRs in its genome. We have studied the transcriptional regulation of these three RNR classes in S. Typhimurium as well as their differential function during infection of macrophage and epithelial cells. Deletion of both NrdR and Fur, two main transcriptional regulators, indicates that Fur specifically represses the class Ib enzyme and that NrdR acts as a global repressor of all three classes. A Fur recognition sequence within the nrdHIEF promoter has also been described and confirmed by electrophoretic mobility shift assays (EMSA). In order to elucidate the role of each RNR class during infection, S. Typhimurium single and double RNR mutants (as well as Fur and NrdR mutants) were used in infection assays with macrophage and epithelial cell lines. Our results indicate class Ia to be mainly responsible for deoxyribonucleotide