The topoisomerase I gene from Candida albicans.
We report here the cloning of the Candida albicans genomic topoisomerase I gene (TOP1) by use of PCR and subsequent hybridization. The predicted protein sequence shared 58.8% identity with the Saccharomyces cerevisiae topoisomerase I and 30-50% identity with other eukaryotic topoisomerase I proteins. A conditional gene disruption strain (CWJ477) was constructed so that one copy of TOP1 was deleted and the other copy of TOP1 was placed under a regulatable promoter. Under repressed conditions, cells grew slowly and cell morphology was abnormal. The virulence of CWJ477 was markedly reduced in a mouse model system, and that of the single gene knockout-strain was slightly attenuated, indicating that TOP1 might play a role in the infection of C. albicans in mice in a dose-dependent manner. Despite the reduced virulence of both the single and double knockout strains, viable cells of the pathogen were recovered from the kidneys as late as 22 d post-infection.
Jiang
Weidong
W
ChemGenics Pharmaceuticals Inc., One Kendall Square, Bldg 300, Cambridge, MA 02139, USA.
Gerhold
David
D
Department of Pharmacology, Jefferson Cancer Institute, Thomas Jefferson University, 233 S. 10th Street, Philadelphia, PA 19107, USA.
Kmiec
Eric B
EB
Department of Pharmacology, Jefferson Cancer Institute, Thomas Jefferson University, 233 S. 10th Street, Philadelphia, PA 19107, USA.
Hauser
Melinda
M
Department of Microbiology and Program in Cellular, Molecular, and Developmental Biology, University of Tennessee, Knoxville, TN 37996-0845, USA.
Becker
Jeffrey M
JM
Department of Microbiology and Program in Cellular, Molecular, and Developmental Biology, University of Tennessee, Knoxville, TN 37996-0845, USA.
Koltin
Yigal
Y
ChemGenics Pharmaceuticals Inc., One Kendall Square, Bldg 300, Cambridge, MA 02139, USA.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002176:Candida albicans / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity*; D003001:Cloning, Molecular; D004264:DNA Topoisomerases, Type I / Q000235:genetics*; D017353:Gene Deletion; D005800:Genes, Fungal; D016681:Genome, Fungal; D007668:Kidney / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D009024:Morphogenesis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D016019:Survival Analysis; D014774:Virulence / Q000235:genetics
D004264:DNA Topoisomerases, Type I