Molecular cloning and characterization of chitinase genes from Candida albicans.

McCreath, Kenneth J.; Specht, Charles A.; Robbins, Phillips W.

doi:10.1073/pnas.92.7.2544

Cited by 93 publications

(100 citation statements)

References 32 publications

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“…Possibly, C. albicans chitinase is O-glycosylated, and underglycosylation favors its release and/or leads to an increased chitinase enzymatic activity in the medium; alternatively, reduced O-glycosylation of cell wall components may impair retention of chitinase in the cell wall. C. albicans chitinase cannot be adsorbed to chitin (41), and specific antibodies are not available, which at present does not allow a more detailed analysis of this phenotype.…”

Section: Resultsmentioning

confidence: 99%

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Multiple Functions of Pmt1p-mediated ProteinO-Mannosylation in the Fungal Pathogen Candida albicans

Timpel¹,

Strahl‐Bolsinger²,

Ziegelbauer³

et al. 1998

Journal of Biological Chemistry

149

186

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Protein mannosylation by Pmt proteins initiates Oglycosylation in fungi. We have identified the PMT1 gene and analyzed the function of Pmt1p in the fungal human pathogen Candida albicans. Mutants defective in PMT1 alleles lacked Pmt in vitro enzymatic activity, showed reduced growth rates, and tended to form cellular aggregates. In addition, multiple specific deficiencies not known in Saccharomyces cerevisiae (including defective hyphal morphogenesis; supersensitivity to the antifungal agents hygromycin B, G418, clotrimazole, and calcofluor white; and reduced adherence to Caco-2 epithelial cells) were observed in pmt1 mutants. PMT1 deficiency also led to faster electrophoretic mobility of the Als1p cell wall protein and to elevated extracellular activities of chitinase. Homozygous pmt1 mutants were avirulent in a mouse model of systemic infection, while heterozygous PMT1/pmt1 strains showed reduced virulence. The results indicate that protein O-mannosylation by Pmt proteins occurs in different fungal species, where PMT1 deficiency can lead to defects in multiple cellular functions.

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Section: Resultsmentioning

confidence: 99%

“…In A, the fragment marked by the asterisks in the schematic diagram (top) was used as probe; in B, a hisG probe was used. (41). To explore if the pmt1 mutation affects the production and/or activity of C. albicans chitinase, we tested its enzymatic activity in the medium and in association with cells (Table III).…”

Section: Resultsmentioning

confidence: 99%

Multiple Functions of Pmt1p-mediated ProteinO-Mannosylation in the Fungal Pathogen Candida albicans

Timpel¹,

Strahl‐Bolsinger²,

Ziegelbauer³

et al. 1998

Journal of Biological Chemistry

149

186

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“…1B, D). The 42 aa repeat was shared by other C. albicans cell wall GPI-modified proteins, including the Cht2 chitinase (Dunkler et al, 2005;McCreath et al, 1995), Hwp1 (Staab & Sundstrom, 1998Sundstrom, 2002), Hwp2 (Sohn et al, 2003, Rbt1 (Braun et al, 2000), Eap1 (Li & Palecek, 2003, Ywp1 (Granger et al, 2005;Sohn et al, 2003), and three putative GPI-modified proteins, Pga6, Pga18 and Pga38. In these proteins the amino acid repeat was 42-46 aa et al, 2007).…”

Section: Pga59 and Pga62 Encode Related Putative Gpimodified Proteinsmentioning

confidence: 99%

The GPI-modified proteins Pga59 and Pga62 of Candida albicans are required for cell wall integrity

et al. 2009

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The fungal cell wall is essential in maintaining cellular integrity and plays key roles in the interplay between fungal pathogens and their hosts. The PGA59 and PGA62 genes encode two short and related glycosylphosphatidylinositol-anchored cell wall proteins and their expression has been previously shown to be strongly upregulated when the human pathogen Candida albicans grows as biofilms. Using GFP fusion proteins, we have shown that Pga59 and Pga62 are cell-walllocated, N-and O-glycosylated proteins. The characterization of C. albicans pga59D/pga59D, pga62D/pga62D and pga59D/pga59D pga62D/pga62D mutants suggested a minor role of these two proteins in hyphal morphogenesis and that they are not critical to biofilm formation. Importantly, the sensitivity to different cell-wall-perturbing agents was altered in these mutants. In particular, simultaneous inactivation of PGA59 and PGA62 resulted in high sensitivity to Calcofluor white, Congo red and nikkomicin Z and in resistance to caspofungin. Furthermore, cell wall composition and observation by transmission electron microscopy indicated an altered cell wall structure in the mutant strains. Collectively, these data suggest that the cell wall proteins Pga59 and Pga62 contribute to cell wall stability and structure. INTRODUCTIONThe cell wall is an essential component of fungal cells, preserving cellular integrity and playing a central role in the interaction of fungi with their environment. This is particularly the case for pathogenic fungi such as the opportunistic yeast pathogen Candida albicans, where the cell wall has been shown to play central roles in adhesion, virulence, biofilm formation, infection and immunomodulation (Albrecht et al., 2006;Douglas, 2003;Netea et al., 2006;Richard et al., 2002b;Sundstrom, 2002). Because of the essential role of the cell wall in cellular integrity and fungal specificity of some enzymes involved in its biogenesis, it is a recognized target for the development of novel antifungals (e.g. echinocandins that target the b-1,3-glucan synthase) (Latge, 2007).The organization of the fungal cell wall has been mainly characterized in the yeasts Saccharomyces cerevisiae and C. albicans and in the filamentous fungus Aspergillus fumigatus (Klis et al., 2006;Latge, 2007;Lesage & Bussey, 2006;Ruiz-Herrera et al., 2006). The yeast cell wall has a bilayered structure. The inner part is composed of a network of b-1,3-glucan molecules linked by hydrogen bonds. These chains can be bound covalently to b-1,6-glucan molecules and to chitin chains. The outer part of the cell wall is composed mainly of mannoproteins (Klis et al., 2006Lesage & Bussey, 2006). Most proteins in the cell wall of ascomycetous yeasts are glycosylphosphatidylinositolanchored proteins (GPI-modified proteins) that become covalently linked to b-1,6-glucan through a remnant of their GPI anchor. As the b-1,6-glucan moiety can be linked to b-1,3-glucan or chitin, the cell wall GPI-modified proteins are strongly linked to the cell wall Klis et al., 2001;Richard & Plaine, 2007).GPI-...

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“…In S. cerevisiae, cell separation requires the controlled action of at least two hydrolytic enzymes: the chitinase encoded by the CTS1 gene (Kuranda and Robbins, 1991) and the endo-␤-1,3-glucanase encoded by ENG1 (Baladrón et al, 2002). In C. albicans, three chitinase genes (CHT1, CHT2 and CHT3) have been cloned and it has been proposed that Cht2p has a similar function to ScCts1p (McCreath et al, 1995). It has also been shown recently that disruption of the ENG1 gene results in the formation of chains of cells (Esteban et al, 2005).…”

Section: Journal Of Cell Science 119 (6)mentioning

confidence: 99%

The Cdc14p phosphatase affects late cell-cycle events and morphogenesis inCandida albicans

Clemente‐Blanco

González-Novo

Machín

et al. 2006

Journal of Cell Science

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We have characterized the CDC14 gene, which encodes a dual-specificity protein phosphatase in Candida albicans, and demonstrated that its deletion results in defects in cell separation, mitotic exit and morphogenesis. The C. albicans cdc14Δ mutants formed large aggregates of cells that resembled those found in ace2-null strains. In cdc14Δ cells, expression of Ace2p target genes was reduced and Ace2p did not accumulate specifically in daughter nuclei. Taken together, these results imply that Cdc14p is required for the activation and daughter-specific nuclear accumulation of Ace2p. Consistent with a role in cell separation, Cdc14p was targeted to the septum region during the M-G1 transition in yeast-form cells. Interestingly, hypha-inducing signals abolished the translocation of Cdc14p to the division plate, and this regulation depended on the cyclin Hgc1p, since hgc1Δ mutants were able to accumulate Cdc14p in the septum region of the germ tubes. In addition to its role in cytokinesis, Cdc14p regulated mitotic exit, since synchronous cultures of cdc14Δ cells exhibited a severe delay in the destruction of the mitotic cyclin Clb2p. Finally, deletion of CDC14 resulted in decreased invasion of solid agar medium and impaired true hyphal growth.

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Molecular cloning and characterization of chitinase genes from Candida albicans.

Abstract: Chitinase (EC 3.2.1.14) is an important enzyme for the remodeling of chitin in the cell wall of fungi. We have cloned three chitinase genes (CHTJ, CHT2, and CHT3)

Cited by 93 publications

References 32 publications

Multiple Functions of Pmt1p-mediated ProteinO-Mannosylation in the Fungal Pathogen Candida albicans

Multiple Functions of Pmt1p-mediated ProteinO-Mannosylation in the Fungal Pathogen Candida albicans

The GPI-modified proteins Pga59 and Pga62 of Candida albicans are required for cell wall integrity

The Cdc14p phosphatase affects late cell-cycle events and morphogenesis inCandida albicans

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