Article Figures & Data
Figures
- Fig. 1.
Strain-specific variations in the HexNAcase activities of C. albicans laboratory strains and clinical isolates (A) and in a set of partially characterized C. albicans clinical isolates (B) (28). HexNAcase activity was measured by the microtiter plate-based assay. The results are the means ± standard deviations of quadruplicate assays.
- Fig. 2.
HexNAcase activities of non-C. albicans Candida species. HexNAcase activity was measured by the microtiter plate-based assay. C. a., Candida albicans (positive control); C. d., Candida dubliniensis; C. r., Candida rugosa. The results are the means ± standard deviations of quadruplicate assays.
- Fig. 3.
Growth and HexNAcase activities of C. albicans clinical isolates I7, I23, and I24. Cells were grown on glucose (25 mM; A and C) or GlcNAc (25 mM; B and D). Cell growth was measured by monitoring the OD600 (A and B). The HexNAcase activities of the culture supernatants were measured at day 7 (C and D). The results are the means ± standard deviations of quadruplicate assays. The strains tested were A72 (positive control; ○, ●), I7 (□, ■), I23 (▵, ▴), and I24 (◊, ⧫).
- Fig. 4.
Southern blot analysis of the HEX1 gene locus in Candida species. Genomic DNA from Candidastrains was restricted with EcoRI, electrophoresed through 0.8% agarose gels, vacuum blotted onto nitrocellulose membranes, and hybridized with a [32P]dCTP-labeled HEX1 DNA probe. Autoradiograms of Southern blots after hybridization under: high-stringency conditions (A) and low-stringency conditions (B) are shown. Lanes: 1, C. albicans A72; 2, C. albicansATCC 10261; 3, C. albicans I7; 4, C. albicansI23; 5, C. albicans I24; 6, C. dubliniensis CD36; 7, C. dubliniensis CD41; 8, C. dubliniensis CD43; 9, C. dubliniensis CD57; 10, C. rugosa TIMM0307; 11, C. rugosa TIMM3489.
Tables
- Table 1.
C. albicans laboratory strains and clinical isolates used in the study
Strain Characteristic or source A72 A reference strain for HexNAcase assay (4, 14); A. Cassone, Instituto Superiore di Sanita, Rome, Italy ATCC 10231 and ATCC 10261 American Type Culture Collection, Manassas, Va. CA2 A. Cassone, Instituto Superiore di Sanita, Rome, Italy CAI4 Δura3::imm434/Δura3::imm434; W. Fonzi, University of California, Irvine C-14-1, C-13-2, D27, D57, D68, D106, D156, D259, D273, and D294 Clinical isolates; School of Dentistry, University of Otago, Dunedin, New Zealand EOB4 HexNAcase-negative mutant (8) IAM12201 Formerly C. stellatoidea; Institute of Applied Microbiology, University of Tokyo, Tokyo, Japan I1 to I47 Clinical isolates; J. Schmid, Massey University, Palmerston North, New Zealand MEN Heterozygous met−, 5FCr; W. L. Whelan, University of Cambridge, Cambridge, United Kingdom R-2477 Clinical isolate; Health Science Center at San Antonio, The University of Texas, San Antonio SGY-243 ade2/ade2, Δura3::ADE2/Δura3::ADE2; R. Kelly, Squibb Institute for Medical Research, Princeton, N.J. TIMM1309 Clinical isolate, formerly C. stellatoidea; Institute of Medical Mycology, Teikyo University, Tokyo, Japan TIMM1369 and TIMM3163 to TIMM3166 Clinical isolates; Institute of Medical Mycology, Teikyo University, Tokyo, Japan 1713D2 Heterozygous HEX1 disruptant (Δhex1::URA3/HEX1); R. D. Cannon, School of Dentistry, University of Otago, Dunedin, New Zealand MN2, MN6, and MN14 to MN16 Clinical isolates (10) MN34 Isolate from healthy individual (10) 93-1591 to 93-2067 Clinical isolates; Health Science Center at San Antonio, The University of Texas, San Antonio - Table 2.
Non-C. albicans Candida, C. neoformans, andS. cerevisiae strains
Species Strain(s) Characteristic or source C. dubliniensis CD36, CD41, CD43, CD57 Clinical isolates; D. C. Coleman, School of Dental Science, University of Dublin, Dublin, Republic of Ireland C. glabrata CBS138 Schimmelcultures, Baarn, The Netherlands CBS2175 Schimmelcultures, Baarn, The Netherlands 850821 ESRa 850920 ESR C. guilliermondii IFO0838 Institute for Fermentation, Osaka, Japan 85.739 ESR 85.791 ESR 89.140 ESR C. kefyr Colindale ESR 78.1161 ESR 78.256 ESR 82.656 ESR B2455 ESR C. krusei IFO0011 Institute for Fermentation, Osaka, Japan 89.102 ESR 89.221 ESR 90.147 ESR B2399 ESR C. lusitaniae TIMM1668 Institute of Medical Mycology, Teikyo University, Tokyo, Japan TIMM3482 Institute of Medical Mycology, Teikyo University, Tokyo, Japan C. parapsilosis MCC499 ESR 90.111 ESR 90.454 ESR 90.463 ESR 90.493 ESR C. rugosa TIMM0307 Institute of Medical Mycology, Teikyo University, Tokyo, Japan TIMM3489 Institute of Medical Mycology, Teikyo University, Tokyo, Japan C. tropicalis IFO0618 Institute for Fermentation, Osaka, Japan 820567 ESR 820738 ESR C. neoformans ATCC 90112 New Zealand Reference Culture Collection, ESR, Wellington, New Zealand (NZRM 3396) ATCC 90113 New Zealand Reference Culture Collection, ESR, Wellington, New Zealand (NZRM 3397) S. cerevisiae AH22 G. R. Fink, Massachussetts Institute of Technology, Cambridge, Mass. (mata, leu2-3, leu2-112, his4-519, can1) y55 J. E. Haber, Brandeis University, Waltham, Mass. (HO, gal3, MAL1, SUC1) 2180A P. A. Sullivan, Massey University, Palmerston North, New Zealand DYC School of Dentistry, University of Otago, Dunedin, New Zealand ↵a ESR, Institute of Environmental Science and Research Health, New Zealand Center for Disease Control, Wellington, New Zealand.
- Table 3.
HexNAcase activities of cell extracts fromCandida species and S. cerevisiaea
Species Strain HexNAcase activity (mmol of pNP min−1 [mg protein]−1) Uninduced (glucose) Induced (GlcNAc) C. albicans A72 0.047 6.66b ATCC 10261 0.023 1.47 SGY-243 0.050 3.82 I1 0.069 3.65 I17 0.060 6.23 I33 0.038 12.46 I44 0.062 5.37 C. dubliniensis CD36 0.029 14.22 CD41 0.031 17.88 C. glabrata CBS138 0 0c C. guilliermondii 85791 0 0 C. krusei B2399 0 0 C. parapsilosis MCC499 0 0 C. tropicalis 820567 0.040 0.149 820738 0.021 0.137 IFO0618 0.059 0.119 S. cerevisiae AH22 0 0c y55 0 0c 2180 0 0c DYC 0 0a ↵a Yeast cells were grown in the presence of either glucose (25 mM; uninduced) or GlcNAc (25 mM; induced) before cell disruption, and the HexNAcase activity was measured by using pNP-GlcNAC as the substrate. The results are the means of two determinations with separate batches of cells, which did not vary by more than 20%.
↵b The value for this enzyme activity is the mean of six determinations, and the standard deviation was ±1.97.
↵c The yeasts did not grow on GlcNAc.
