The nucleotide (nt) sequence of the dihydrofolate reductase (DHFR)-encoding gene (DFR1) of Candida albicans was determined. The gene contains an open reading frame of 576 nt, coding for a protein of 192 amino acid (aa) residues (calculated M(r) 22,222), that is 38.5 and 31% similar to the Saccharomyces cerevisiae and human enzymes, respectively. The first 36 residues, at the N terminus, of the deduced aa sequence are identical to those determined by sequencing of the purified enzyme from C. albicans. Putative transcription start points were also determined. Restriction-fragment-length polymorphism analysis of the DFR1 chromosomal region suggests the presence of a single copy of the gene per haploid genome and shows a limited variability among the different C. albicans strains tested.
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http://dx.doi.org/10.1016/0378-1119(94)90049-3 | DOI Listing |
Insects
December 2024
Department of Entomology, University of Minnesota, St. Paul, MN 55108, USA.
Bacterial and eukaryotic dihydrofolate reductase (DHFR) enzymes are essential for DNA synthesis and are differentially sensitive to the competitive inhibitors trimethoprim and methotrexate. Unexpectedly, trimethoprim did not reduce abundance, and the Stri DHFR homolog contained amino acid substitutions associated with trimethoprim resistance in . A phylogenetic tree showed good association of DHFR protein sequences with supergroup A and B assignments.
View Article and Find Full Text PDFPrep Biochem Biotechnol
January 2025
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Chinese hamster ovary (CHO) cells represent the most common host system for the expression of high-quality recombinant proteins. The development of stable CHO cell lines used in industrial recombinant protein production often relies on dihydrofolate reductase (DHFR) and glutamine synthetase (GS) amplification systems. Conventional approaches to develop stable cell lines lead to heterogeneous cell populations.
View Article and Find Full Text PDFInt J Biol Macromol
January 2025
Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand; Center of Excellence in Molecular Crop, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand. Electronic address:
A dihydrofolate reductase (DHFR)-like enzyme from Leptospira interrogans (LiDHFRL) was cloned and the recombinant protein was characterized. Sequence alignment suggested that the enzyme lacked the conserved catalytic residues found in DHFR. Indeed, LiDHFRL did not catalyze the reduction of dihydrofolate by either NADH or NADPH.
View Article and Find Full Text PDFStructure
December 2024
Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, 2200 Copenhagen, Denmark. Electronic address:
Gene variants resulting in insertions or deletions of amino acid residues (indels) have important consequences for evolution and are often linked to disease, yet, compared to missense variants, the effects of indels are poorly understood and predicted. We developed a sensitive protein folding sensor based on the complementation of uracil auxotrophy in yeast by circular permutated orotate phosphoribosyltransferase (CPOP). The sensor reports on the folding of disease-linked missense variants and de-novo-designed proteins.
View Article and Find Full Text PDFNat Commun
November 2024
Institute for Global Health and Infectious Diseases, University of North Carolina, Chapel Hill, NC, USA.
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