Cloning and characterization of ribonucleotide reductase from Chlamydia trachomatis

64Citations
Citations of this article
31Readers
Mendeley users who have this article in their library.

This article is free to access.

Abstract

In all organisms the deoxyribonucleotide precursors required for DNA synthesis are synthesized from ribonucleotides, a reaction catalyzed by ribonucleotide reductase. In a previous study we showed that Chlamydia trachomatis growth was inhibited by hydroxyurea, an inhibitor of ribonucleotide reductase, and a mutant resistant to the cytotoxic effects of the drug was isolated. Here we report the cloning, expression, and purification of the R1 and R2 subunits of the C. trachomatis ribonucleotide reductase. In comparison with other ribonucleotide reductases, the primary sequence of protein R1 has an extended amino terminus, and the R2 protein has a phenylalanine where the essential tyrosine is norreally located. Despite its unusual primary structure, the recombinant enzyme catalyzes the reduction of CDP to dCDP. Results from deletion mutagenesis experiments indicate that while the extended amino terminus of the R1 protein is not required for enzyme activity, it is needed for allosteric inhibition mediated by dATP. Resuits with site-directed mutants of protein R2 suggest that the essential tyrosine is situated two amino acids downstream of its normal location. Finally, Western blot analysis show that the hydroxyurea-resistant mutant C. trachomatis isolate overexpresses both subunits of ribonucleotide reductase. At the genetic level, compared with wild type C. trachomatis, the resistant isolate has a single base mutation just upstream of the ATG start cedon of the R2 protein. The possibility that this mutation affects translational efficiency is discussed.

References Powered by Scopus

The 3' terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites

2689Citations
N/AReaders
Get full text

Genome sequence of an obligate intracellular pathogen of humans: Chlamydia trachomatis

1358Citations
N/AReaders
Get full text

Reduction of ribonucleotides.

952Citations
N/AReaders
Get full text

Cited by Powered by Scopus

Ribonucleotide reductases

909Citations
N/AReaders
Get full text

Structure, function, and mechanism of ribonucleotide reductases

378Citations
N/AReaders
Get full text

Zoonotic Chlamydophila psittaci infections from a clinical perspective

224Citations
N/AReaders
Get full text

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Cite

CITATION STYLE

APA

Roshick, C., Iliffe-Lee, E. R., & McClarty, G. (2000). Cloning and characterization of ribonucleotide reductase from Chlamydia trachomatis. Journal of Biological Chemistry, 275(48), 38111–38119. https://doi.org/10.1074/jbc.M006367200

Readers' Seniority

Tooltip

PhD / Post grad / Masters / Doc 8

35%

Researcher 8

35%

Professor / Associate Prof. 4

17%

Lecturer / Post doc 3

13%

Readers' Discipline

Tooltip

Agricultural and Biological Sciences 10

42%

Chemistry 6

25%

Biochemistry, Genetics and Molecular Bi... 6

25%

Immunology and Microbiology 2

8%

Save time finding and organizing research with Mendeley

Sign up for free