This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Dallas, W S Articles by Dev, I K Search for Related Content PubMed PubMed Citation Articles by Dallas, W S Articles by Dev, I K

research-article

Cloning, sequencing, and enhanced expression of the dihydropteroate synthase gene of Escherichia coli MC4100.

Department of Molecular Genetics and Microbiology, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.

ABSTRACT

The Escherichia coli gene coding for dihydropteroate synthase (DHPS) has been cloned and sequenced. The protein has 282 amino acids and a compositional molecular mass of 30,314 daltons. Increased expression of the enzyme was realized by using a T7 expression system. The enzyme was purified and crystallized. A temperature-sensitive mutant was isolated and found to express a DHPS with a lower specific activity and lower affinities for para-aminobenzoic acid and sulfathiazole. The allele had a point mutation that changed a phenylalanine codon to a leucine codon, and the mutation was in a codon that is conserved among published DHPS sequences.

This article has been cited by other articles:

• Levin, I., Mevarech, M., Palfey, B. A. (2007). Characterization of a Novel Bifunctional Dihydropteroate Synthase/Dihydropteroate Reductase Enzyme from Helicobacter pylori. J. Bacteriol. 189: 4062-4069 [Abstract] [Full Text]  
• Scollard, D. M., Adams, L. B., Gillis, T. P., Krahenbuhl, J. L., Truman, R. W., Williams, D. L. (2006). The Continuing Challenges of Leprosy. Clin. Microbiol. Rev. 19: 338-381 [Abstract] [Full Text]  
• Gregson, A., Plowe, C. V. (2005). Mechanisms of Resistance of Malaria Parasites to Antifolates. Pharmacol. Rev. 57: 117-145 [Abstract] [Full Text]  
• Guldener, U., Koehler, G. J., Haussmann, C., Bacher, A., Kricke, J., Becher, D., Hegemann, J. H. (2004). Characterization of the Saccharomyces cerevisiae Fol1 Protein: Starvation for C1 Carrier Induces Pseudohyphal Growth. Mol. Biol. Cell 15: 3811-3828 [Abstract] [Full Text]  
• Zingale, A., Carrera, P., Lazzarin, A., Scarpellini, P. (2003). Detection of Pneumocystis carinii and Characterization of Mutations Associated with Sulfa Resistance in Bronchoalveolar Lavage Samples from Human Immunodeficiency Virus-Infected Subjects. J. Clin. Microbiol. 41: 2709-2712 [Abstract] [Full Text]  
• Hayton, K., Ranford-Cartwright, L. C., Walliker, D. (2002). Sulfadoxine-Pyrimethamine Resistance in the Rodent Malaria Parasite Plasmodium chabaudi. Antimicrob. Agents Chemother. 46: 2482-2489 [Abstract] [Full Text]  
• Williams, D. L., Pittman, T. L., Gillis, T. P., Matsuoka, M., Kashiwabara, Y. (2001). Simultaneous Detection of Mycobacterium leprae and Its Susceptibility to Dapsone Using DNA Heteroduplex Analysis. J. Clin. Microbiol. 39: 2083-2088 [Abstract] [Full Text]  
• Takahashi, T., Hosoya, N., Endo, T., Nakamura, T., Sakashita, H., Kimura, K., Ohnishi, K., Nakamura, Y., Iwamoto, A. (2000). Relationship between Mutations in Dihydropteroate Synthase of Pneumocystis carinii f. sp. hominis Isolates in Japan and Resistance to Sulfonamide Therapy. J. Clin. Microbiol. 38: 3161-3164 [Abstract] [Full Text]  
• Williams, D. L., Spring, L., Harris, E., Roche, P., Gillis, T. P. (2000). Dihydropteroate Synthase of Mycobacterium leprae and Dapsone Resistance. Antimicrob. Agents Chemother. 44: 1530-1537 [Abstract] [Full Text]  
• Qvarnström, Y., Swedberg, G. (2000). Additive effects of a two-amino-acid insertion and a single-amino-acid substitution in dihydropteroate synthase for the development of sulphonamide-resistant Neisseria meningitidis. Microbiology 146: 1151-1156 [Abstract] [Full Text]  
• Nopponpunth, V., Sirawaraporn, W., Greene, P. J., Santi, D. V. (1999). Cloning and Expression of Mycobacterium tuberculosis and Mycobacterium leprae Dihydropteroate Synthase in Escherichia coli. J. Bacteriol. 181: 6814-6821 [Abstract] [Full Text]  
• Gibreel, A., Sköld, O. (1999). Sulfonamide Resistance in Clinical Isolates of Campylobacter jejuni: Mutational Changes in the Chromosomal Dihydropteroate Synthase. Antimicrob. Agents Chemother. 43: 2156-2160 [Abstract] [Full Text]  
• Padayachee, T., Klugman, K. P. (1999). Novel Expansions of the Gene Encoding Dihydropteroate Synthase in Trimethoprim-Sulfamethoxazole-Resistant Streptococcus pneumoniae. Antimicrob. Agents Chemother. 43: 2225-2230 [Abstract] [Full Text]  
• Berlyn, M. K. B. (1998). Linkage Map of Escherichia coli K-12, Edition 10: The Traditional Map. Microbiol. Mol. Biol. Rev. 62: 814-984 [Abstract] [Full Text]  
• Swedberg, G., Ringertz, S., Sköld, O. (1998). Sulfonamide Resistance in Streptococcus pyogenes Is Associated with Differences in the Amino Acid Sequence of Its Chromosomal Dihydropteroate Synthase. Antimicrob. Agents Chemother. 42: 1062-1067 [Abstract] [Full Text]  
• Chen, W.-J., Jayawickreme, C., Watson, C., Wolfe, L., Holmes, W., Ferris, R., Armour, S., Dallas, W., Chen, G., Boone, L., Luther, M., Kenakin, T. (1998). Recombinant Human CXC-Chemokine Receptor-4 in Melanophores Are Linked to Gi Protein: Seven Transmembrane Coreceptors for Human Immunodeficiency Virus Entry into Cells. Mol. Pharmacol. 53: 177-181 [Abstract] [Full Text]  
• Vedantam, G., Guay, G. G., Austria, N. E., Doktor, S. Z., Nichols, B. P. (1998). Characterization of Mutations Contributing to Sulfathiazole Resistance in Escherichia coli. Antimicrob. Agents Chemother. 42: 88-93 [Abstract] [Full Text]  
• Triglia, T., Menting, J. G. T., Wilson, C., Cowman, A. F. (1997). Mutations in dihydropteroate synthase are responsible for sulfone and sulfonamide resistance in falciparum. Proc. Natl. Acad. Sci. USA 94: 13944-13949 [Abstract] [Full Text]  
• Neuburger, M., Rébeillé, F., Jourdain, A.ès, Nakamura, S., Douce, R. (1996). Mitochondria Are a Major Site for Folate and Thymidylate Synthesis in Plants. J. Biol. Chem. 271: 9466-9472 [Abstract] [Full Text]  
• Mengin-Lecreulx, D., van Heijenoort, J. (1996). Characterization of the Essential Gene glmM Encoding Phosphoglucosamine Mutase in Escherichia coli. J. Biol. Chem. 271: 32-39 [Abstract] [Full Text]