The polymorphism of protein phosphatase Z1 gene in Candida albicans

Abstract

The gene of protein phosphatase Z1 (CaPPZ1) that codes a fungus specific regulatory enzyme was investigated in Candida albicans. After cloning and sequencing CaPPZ1 we revealed the heterozygous nature of the ATCC 10231 reference strain, and identified two new alleles termed CaPPZ1-2 and CaPPZ1-3. The genetic polymorphism in CaPPZ1 was extended by finding a fourth allele CaPPZ1-4 in a clinical isolate. Single nucleotide replacements and short in/del mutations were identified in the gene, some of which resulted in amino acid changes in the protein. The analysis of the hypervariable 3'-noncoding gene region in 27 DNA sequences obtained from reference strains and clinical samples confirmed the presence of four distinct DNA sequence-groups that correspond to the four main alleles of CaPPZ1. In addition to the allelic combinations, we detected individual mutations elevating genetic variability of the opportunistic pathogen. We utilized the hypervariable gene region for genotyping C. albicans in clinical isolates by sequencing the cloned amplified region, by direct sequencing of the PCR products, or by RFLP analysis. The comparison of the genotypes of the strains originating from different body parts of the same patient proved to be useful in delineating the origin of the infection.

Molecular Insights into the Fungus-Specific Serine/Threonine Protein Phosphatase Z1 in Candida albicans

“…GQ357913 [14, 36]) and cloned into the E. coli expression vector pGEX6p-1 (Amersham Biosciences); both constructs were sequence verified (UD Genomed, Ltd.). Both constructs were then transformed into E. coli BL21(DE3) RIL cells (Agilent), and expression of N-terminal glutathione S -transferase (GST)-tagged proteins was induced with 0.6 mM IPTG (Sigma) at 18°C.…”

The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase: A mutagenesis analysis

“…The fact that Ppz1 is found only in fungi, including pathogenic ones, such as Candida albicans 15 , 23 or Aspergillus fumigatus 24 and that increased Ppz1 function has a detrimental effect on yeast growth 9 , 10 , 13 suggest that this enzyme may represent an interesting target for the development of antifungal therapies. However, because of the relative similarity between the catalytic domain of Ppz1 and that of the ubiquitous type 1 enzymes, it might be argued that specificity could be an important issue when interfering with Ppz1 function in vivo .…”

The protein phosphatase PhzA of A. fumigatus is involved in oxidative stress tolerance and fungal virulence

“…These phosphatases are composed of a highly conserved COOH-terminal catalytic domain similar to the type 1 S/T phosphatases while the unstructured NH2-terminal regulatory domain contains a N-myristoylation site and a Arg/Ser rich motif that are unique to these fungal proteins (Arino, 2002; Leiter et al, 2012; Posas et al, 1993). Ppz proteins have been extensively characterized in Saccharomyces cerevisiae , Schizzosaccharomces pombe and Candida albicans and shown to be involved in salt tolerance, cell wall integrity, cell cycle regulation, and oxidative stress tolerance (Balcells et al, 1997; Kovacs et al, 2010; Posas et al, 1993, 1995). However, although the Neurospora crassa or Aspergillus nidulans and Aspergillus fumigatus PPZ are able to complement Δ ppz deletion mutants of S. cerevisiae , Ppz proteins of filamentous fungi were shown to be exclusively involved in resistance to oxidative stress (Leiter et al, 2012; Vissi et al, 2001).…”