Isolation of aCandida albicans gene, tightly linked toURA3, coding for a putative transcription factor that suppresses aSaccharomyces cerevisiaeaft1 mutation

García, Micaela Gómez; O’Connor, José-Enrique; García, Lorena Latorre; Martínez, Sami Irar; Herrero, Enrique; Agudo, Lucas del Castillo

doi:10.1002/1097-0061(20010315)18:4<301::aid-yea672>3.0.co;2-h

Cited by 38 publications

(4 citation statements)

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“…Expression of IRO1 in a Δ aft1 S. cerevisiae strain restored growth in iron depleted media. However, a role of IRO1 in C. albicans iron metabolism is not confirmed [56]. On the other hand, ectopic expression of URA3 has been shown to affect several features of C. albicans , such as hyphal morphology, adhesion, virulence and cellular proteome in addition to Ura3p activity [57,58].…”

Section: Discussionmentioning

confidence: 99%

Involvement of the mitogen activated protein kinase Hog1p in the response of Candida albicansto iron availability

et al. 2013

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BackgroundIron is an essential nutrient for almost all organisms, and generating iron limiting conditions for pathogens is one of the host defense strategies against microbial infections. Excess of iron can be toxic; therefore, iron uptake is tightly controlled. The high affinity iron uptake system of the opportunistic pathogenic yeast Candida albicans has been shown to be essential for virulence. Several transcription factors and regulators of iron uptake genes were identified, but the knowledge of signaling pathways is still limited. Gene expression profiling of the Δhog1 deletion mutant indicated an involvement of the mitogen activated protein (MAP) kinase Hog1p. However, the function of Hog1p in the response of C. albicans to iron availability was not studied in detail. Thus, we analyzed phenotypic and molecular responses of C. albicans to different iron concentrations particularly with respect to the activity of the Hog1p MAP kinase module.ResultsWe observed flocculation of yeast cells, when the iron ion concentration was equal to or higher than 5 μM. This phenotype was dependent on the MAP kinase Hog1p and the corresponding MAP kinase kinase Pbs2p. Moreover, high extracellular iron ion concentrations led to hyper-phosphorylation of Hog1p. We determined lower amounts of multicopper ferroxidase (MCFO) proteins and lower ferric reductase activity, when the iron ion concentration in the medium was increased. This effect was also observed for the Δhog1 mutant. However, the amounts of MCFO proteins and the cell surface ferric reductase activity were increased in the Δhog1 in comparison to wild type cells. This effect was independent of iron availability in growth media.ConclusionsIn C. albicans, the MAP kinase Hog1p is part of the network regulating the response of the organism to iron availability. Hog1p was transiently phosphorylated under high iron concentrations and was essential for a flocculent phenotype. Furthermore, deletion of HOG1 led to increased levels of components of the reductive iron uptake system in comparison to the wild-type, independent of iron concentrations in the media. However, the additional induction of this system by low iron concentrations was independent of HOG1.

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Section: Discussionmentioning

confidence: 99%

Involvement of the mitogen activated protein kinase Hog1p in the response of Candida albicansto iron availability

et al. 2013

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“…When it is successful, the construct still leaves behind a sizable hisG sequence at the target site, which could impact adjacent gene expression or be involved in further gene rearrangement events. Recombination between hisG sequences can also be imprecise, leading to the deletion of neighboring sequences (García et al, 2001 ). Choosing to express URA3 at an ectopic site can also reduce the virulence of the strain and result in a phenotype that is mistakenly associated with the mutation of the gene at the target locus (Alani et al, 1987 ; Lay et al, 1998 ; Cheng et al, 2003 ; Brand et al, 2004 ; Samaranayake and Hanes, 2011 ).…”

Section: Genetic Manipulation Of Candida Species: Historical Contextmentioning

confidence: 99%

CRISPR-Based Genetic Manipulation of Candida Species: Historical Perspectives and Current Approaches

et al. 2021

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The Candida genus encompasses a diverse group of ascomycete fungi that have captured the attention of the scientific community, due to both their role in pathogenesis and emerging applications in biotechnology; the development of gene editing tools such as CRISPR, to analyze fungal genetics and perform functional genomic studies in these organisms, is essential to fully understand and exploit this genus, to further advance antifungal drug discovery and industrial value. However, genetic manipulation of Candida species has been met with several distinctive barriers to progress, such as unconventional codon usage in some species, as well as the absence of a complete sexual cycle in its diploid members. Despite these challenges, the last few decades have witnessed an expansion of the Candida genetic toolbox, allowing for diverse genome editing applications that range from introducing a single point mutation to generating large-scale mutant libraries for functional genomic studies. Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology is among the most recent of these advancements, bringing unparalleled versatility and precision to genetic manipulation of Candida species. Since its initial applications in Candida albicans, CRISPR-Cas9 platforms are rapidly evolving to permit efficient gene editing in other members of the genus. The technology has proven useful in elucidating the pathogenesis and host-pathogen interactions of medically relevant Candida species, and has led to novel insights on antifungal drug susceptibility and resistance, as well as innovative treatment strategies. CRISPR-Cas9 tools have also been exploited to uncover potential applications of Candida species in industrial contexts. This review is intended to provide a historical overview of genetic approaches used to study the Candida genus and to discuss the state of the art of CRISPR-based genetic manipulation of Candida species, highlighting its contributions to deciphering the biology of this genus, as well as providing perspectives for the future of Candida genetics.

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“…First, a portion of the 3′ end of the IRO1 involving in iron utilization was removed during the construction of the parental strain CAI4 (Ura − ), which activated the alternative mechanisms of iron uptake in CAI4. The growth of CAI4 is better than that of the wild-type SC5314 strain in an iron-restricted environment 27 - 29 . Second, one copy of hisG sequences left in the chromosome decreases the integration frequency of the disruption cassette at the target locus because of the competition between the endogenous hisG sequences and the successive hisG cassettes in the second round of transformation 30 , 31 .…”

Section: Cassettes Used In Gene Disruptionmentioning

confidence: 99%

Molecular genetic techniques for gene manipulation inCandida albicans

Yan

et al. 2014

Virulence

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Candida albicans is one of the most common fungal pathogen in humans due to its high frequency as an opportunistic and pathogenic fungus causing superficial as well as invasive infections in immunocompromised patients. An understanding of gene function in C. albicans is necessary to study the molecular basis of its pathogenesis, virulence and drug resistance. Several manipulation techniques have been used for investigation of gene function in C. albicans, including gene disruption, controlled gene expression, protein tagging, gene reintegration, and overexpression. In this review, the main cassettes containing selectable markers used for gene manipulation in C. albicans are summarized; the advantages and limitations of these cassettes are discussed concerning the influences on the target gene expression and the virulence of the mutant strains.

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Isolation of aCandida albicans gene, tightly linked toURA3, coding for a putative transcription factor that suppresses aSaccharomyces cerevisiaeaft1 mutation

Cited by 38 publications

References 56 publications

Involvement of the mitogen activated protein kinase Hog1p in the response of Candida albicansto iron availability

Involvement of the mitogen activated protein kinase Hog1p in the response of Candida albicansto iron availability

CRISPR-Based Genetic Manipulation of Candida Species: Historical Perspectives and Current Approaches

Molecular genetic techniques for gene manipulation inCandida albicans

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