FLO8 deletion leads to decreased adhesion and virulence with downregulated expression of EPA1, EPA6, and EPA7 in Candida glabrata

Zhao, Juntao; Chen, Ke-Zhi; Liu, Jinyan; Li, Weihua; Wang, Yuzhu; Wang, Lu-Ling; Xiang, Ming-Jie

doi:10.1007/s42770-022-00703-7

Cited by 5 publications

(8 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To knockout the Cdc50 gene in C. glabrata , an NAT cassette containing a nourseothricin resistance marker was used for homologous recombination, as described previously [ 55 ]. Briefly, the upstream and downstream regions of Cdc50 were amplified using the DNA of strain ATCC2001 as the template and primers Cdc50-UP and Cdc50-DOWN, and the NAT marker was amplified using plasmid pYC44 and primers NAT-Cdc50 (the reverse complement sequences are marked in lower case in Table 4 ).…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, the upstream and downstream regions of Cdc50 were amplified using the DNA of strain ATCC2001 as the template and primers Cdc50-UP and Cdc50-DOWN, and the NAT marker was amplified using plasmid pYC44 and primers NAT-Cdc50 (the reverse complement sequences are marked in lower case in Table 4 ). After fusion PCR, a knockout cassette was transformed into the ATCC2001 strain, as described previously by Zhao et al [ 55 ]. Δcdc50 transformants that could grow on plates containing 100 μg/ml nourseothricin were selected and their genotypes were confirmed using PCR with primers y-incdc50 and y-outcdc50.…”

Section: Methodsmentioning

confidence: 99%

“…To investigate the virulence of different C. glabrata strains, the infection model organism Caenorhabditis elegans was used, with Escherichia coli strain OP50 as a quality control. The protocol followed the instructions of Zhao et al [ 55 ]. Briefly, after washing with M9 buffer three times, C. elegans (strain glp-4(bn2) I) at the L4 stage were transferred to brain heart infusion (BHI) medium plates containing yeast or to nematode growth medium (NGM) plates containing OP50 cells.…”

Section: Methodsmentioning

confidence: 99%

“…The approach used for qRT-PCR followed the method of Zhao et al [ 55 ]. In short, C. glabrata cells in mid-log phase were collected, washed and frozen; then total RNA was isolated using a yeast RNAiso kit (Takara), followed by treatment with gRNA Eraser (Takara) to remove residual genomic DNA.…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

P4-ATPase subunit Cdc50 plays a role in yeast budding and cell wall integrity in Candida glabrata

et al. 2023

Self Cite

View full text Add to dashboard Cite

Background As highly-conserved types of lipid flippases among fungi, P4-ATPases play a significant role in various cellular processes. Cdc50 acts as the regulatory subunit of flippases, forming heterodimers with Drs2 to translocate aminophospholipids. Cdc50 homologs have been reported to be implicated in protein trafficking, drug susceptibility, and virulence in Saccharomyces cerevisiae, Candida albicans and Cryptococcus neoformans. It is likely that Cdc50 has an extensive influence on fungal cellular processes. The present study aimed to determine the function of Cdc50 in Candida glabrata by constructing a Δcdc50 null mutant and its complemented strain. Results In Candida glabrata, the loss of Cdc50 led to difficulty in yeast budding, probably caused by actin depolarization. The Δcdc50 mutant also showed hypersensitivity to azoles, caspofungin, and cell wall stressors. Further experiments indicated hyperactivation of the cell wall integrity pathway in the Δcdc50 mutant, which elevated the major cell wall contents. An increase in exposure of β-(1,3)-glucan and chitin on the cell surface was also observed through flow cytometry. Interestingly, we observed a decrease in the phagocytosis rate when the Δcdc50 mutant was co-incubated with THP-1 macrophages. The Δcdc50 mutant also exhibited weakened virulence in nematode survival tests. Conclusion The results suggested that the lipid flippase subunit Cdc50 is implicated in yeast budding and cell wall integrity in C. glabrata, and thus have a broad influence on drug susceptibility and virulence. This work highlights the importance of lipid flippase, and offers potential targets for new drug research.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

P4-ATPase subunit Cdc50 plays a role in yeast budding and cell wall integrity in Candida glabrata

et al. 2023

Self Cite

View full text Add to dashboard Cite

show abstract

“…Comparative analysis of VdPBP1 gene expression in both standard MS medium and the MS environment within tomato plants revealed a significant upregulation in its expression. Additionally, it is known that the ORF harboring 313 amino acids at the protein’s C-terminal can restore adhesion in the FLO8 mutant [ 3 ], a transcription factor responsible for regulating the expression of flocculation genes in S. cerevisiae [ 8 , 9 ]. This indicates a potential role for the VdPBP1 gene in early host infection stages, such as root attachment and resistance to root toxins.…”

Section: Introductionmentioning

confidence: 99%

Verticillium dahliae VdPBP1 Transcription Factor Is Required for Hyphal Growth, Virulence, and Microsclerotia Formation

Nguyen,

Duong,

Nguyen

et al. 2024

Microorganisms

View full text Add to dashboard Cite

Verticillium dahliae, a fungal pathogen that affects more than 200 plant species, including tomatoes, requires specific proteins for its early steps in plant infection. One such crucial protein, VdPBP1, exhibits high expression in the presence of tomato roots. Its 313-amino acid C-terminal section restores adhesion in nonadhesive Saccharomyces cerevisiae strains. To uncover its role, we employed a combination of bioinformatics, genetics, and morphological analyses. Our findings underscore the importance of VdPBP1 in fungal growth and pathogenesis. Bioinformatic analysis revealed that the VdPBP1 gene consists of four exons and three introns, encoding a 952-codon reading frame. The protein features a 9aaTAD domain, LsmAD, and PAB1 DNA-binding sites, as well as potential nuclear localization and transmembrane helix signals. Notably, the deletion of a 1.1 kb fragment at the gene’s third end impedes microsclerotia formation and reduces pathogenicity. Mutants exhibit reduced growth and slower aerial mycelial development compared to the wild type. The VdPBP1 deletion strain does not induce disease symptoms in tomato plants. Furthermore, VdPBP1 deletion correlates with downregulated microsclerotia formation-related genes, and promoter analysis reveals regulatory elements, including sites for Rfx1, Mig1, and Ste12 proteins. Understanding the regulation and target genes of VdPBP1 holds promise for managing Verticillium wilt disease and related fungal pathogens.

show abstract

Biofilm Formation in Candida Species

Elibe,

Innocent

2024

Recent Advances in Human Fungal Diseases

View full text Add to dashboard Cite

FLO8 deletion leads to decreased adhesion and virulence with downregulated expression of EPA1, EPA6, and EPA7 in Candida glabrata

Cited by 5 publications

References 40 publications

P4-ATPase subunit Cdc50 plays a role in yeast budding and cell wall integrity in Candida glabrata

P4-ATPase subunit Cdc50 plays a role in yeast budding and cell wall integrity in Candida glabrata

Verticillium dahliae VdPBP1 Transcription Factor Is Required for Hyphal Growth, Virulence, and Microsclerotia Formation

Biofilm Formation in Candida Species

Contact Info

Product

Resources

About