A NovelcisElement Achieves the Same Solution as an AncestralcisElement During Thiamine Starvation inCandida glabrata

Iosue, Christine L.; Gulotta, Anthony P; Selhorst, Kathleen B; Mody, Alison C; Barbour, Kristin M; Marcotte, Meredith J; Bui, Lilian N; Leone, Sarah G; Lang, Emma C.; Hughes, Genevieve H; Wykoff, Dennis D.

doi:10.1534/g3.119.400897

Cited by 3 publications

(11 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…glabrata THI promoters, raising the question of how a promoter that does not have the same cis architecture as other THI promoters is still responsive to the same trans environment in S . cerevisiae [ 7 ]. CgPMU3 is a recently evolved phosphatase gene that cleaves thiamine pyrophosphate (TPP), is required for C .…”

Section: Resultsmentioning

confidence: 99%

“…CgPMU3 having a different critical cis element relative to other promoters suggests that the architecture of the CgPMU3 promoter is different from other THI promoters, even though CgPMU3 uses both Pdc2 and Thi3 to upregulate expression during thiamine starvation [ 7 ]. We hypothesized that CgPMU3 has a different sensitivity to Pdc2 relative to other THI promoters, either not requiring all of Pdc2, or being differentially sensitive to parts of Pdc2.…”

Section: Resultsmentioning

confidence: 99%

“…cerevisiae . To do this, we cloned 30 bp to 100 bp fragments around the cis elements [ 7 ] known to be important for regulation into the CgPMU1 promoter ( Fig 8A ). We identified regions of varying lengths that are sufficient for conferring thiamine regulation to the CgPMU1 promoter: 90 bp of the ScPDC5 promoter, 60 bp of the CgTHI20 promoter, and 100 bp of the CgPMU3 promoter ( Fig 8B ).…”

Section: Resultsmentioning

confidence: 99%

“…cerevisiae wild-type and C . glabrata wild-type, as well as strains in which the thiamine pathway regulators were deleted: Scpdc2Δ , Scthi3Δ , Scthi2Δ , Cgpdc2Δ , and Cgthi3Δ [ 6 , 7 , 17 – 19 ].…”

Section: Methodsmentioning

confidence: 99%

“…Nosaka et al identified an upstream region of PDC5 pr between -416 and -346 nucleotides that is necessary for ScPDC5 expression, and this region shares sequence similarity with a binding site in THI promoters that we identified in C . glabrata [ 7 , 12 ]. Because deletion of ScPDC2 is lethal, few genome-wide studies have examined Pdc2 properties in S .…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Pyruvate decarboxylase and thiamine biosynthetic genes are regulated differently by Pdc2 in S. cerevisiae and C. glabrata

et al. 2023

Self Cite

View full text Add to dashboard Cite

Understanding metabolism in the pathogen Candida glabrata is key to identifying new targets for antifungals. The thiamine biosynthetic (THI) pathway is partially defective in C. glabrata, but the transcription factor CgPdc2 upregulates some thiamine biosynthetic and transport genes. One of these genes encodes a recently evolved thiamine pyrophosphatase (CgPMU3) that is critical for accessing external thiamine. Here, we demonstrate that CgPdc2 primarily regulates THI genes. In Saccharomyces cerevisiae, Pdc2 regulates both THI and pyruvate decarboxylase (PDC) genes, with PDC proteins being a major thiamine sink. Deletion of PDC2 is lethal in S. cerevisiae in standard growth conditions, but not in C. glabrata. We uncover cryptic cis elements in C. glabrata PDC promoters that still allow for regulation by ScPdc2, even when that regulation is not apparent in C. glabrata. C. glabrata lacks Thi2, and it is likely that inclusion of Thi2 into transcriptional regulation in S. cerevisiae allows for a more complex regulation pattern and regulation of THI and PDC genes. We present evidence that Pdc2 functions independent of Thi2 and Thi3 in both species. The C-terminal activation domain of Pdc2 is intrinsically disordered and critical for species differences. Truncation of the disordered domains leads to a gradual loss of activity. Through a series of cross species complementation assays of transcription, we suggest that there are multiple Pdc2-containing complexes, and C. glabrata appears to have the simplest requirement set for THI genes, except for CgPMU3. CgPMU3 has different cis requirements, but still requires Pdc2 and Thi3 to be upregulated by thiamine starvation. We identify the minimal region sufficient for thiamine regulation in CgTHI20, CgPMU3, and ScPDC5 promoters. Defining the cis and trans requirements for THI promoters should lead to an understanding of how to interrupt their upregulation and provide targets in metabolism for antifungals.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Pyruvate decarboxylase and thiamine biosynthetic genes are regulated differently by Pdc2 in S. cerevisiae and C. glabrata

et al. 2023

Self Cite

View full text Add to dashboard Cite

show abstract

The cloned SPI-1 type 3 secretion system can be functionally expressed outside Salmonella backgrounds

Patel

Cangelosi

Warrier

et al. 2020

FEMS Microbiology Letters

View full text Add to dashboard Cite

Due to its potential for use in bacterial engineering applications, we previously cloned the SPI-1 type 3 secretion system (T3SS) genes from the genome of Salmonella enterica serovar Typhimurium strain LT2. We have documented that this clone, while functionally expressed in S. Typhimurium strains, displays a severe expression defect in other Gram negative backgrounds including Escherichia coli. To address this issue, we compared SPI-1 DNA sequence across different backgrounds, fully sequenced the original SPI-1 clone, and cloned SPI-1 from other S. Typhimurium strains. In this process, we were able to successfully obtain SPI-1 clones that are functionally expressed in E. coli indicating the first such result for a full-length SP-1 T3SS clone. We discovered that the original cloning technique using a DNA homology-based capture method was the root of the expression defect and that the FRT-Capture technique is preferable over the homology-based method. This result paves the way for future studies and applications using cloned SPI-1 and other T3SS in non-Salmonella bacterial backgrounds.

show abstract

Regulation of thiamine and pyruvate decarboxylase genes by Pdc2 in Nakaseomyces glabratus (Candida glabrata) is complex

Dottor,

Iosue,

Loshnowsky

et al. 2024

G3: Genes, Genomes, Genetics

View full text Add to dashboard Cite

Thiamine (vitamin B1) is essential for glucose catabolism. In the yeast species, Nakaseomyces glabratus (formerly Candida glabrata) and Saccharomyces cerevisiae, the transcription factor Pdc2 (with Thi3 and Thi2) upregulates pyruvate decarboxylase (PDC) genes and thiamine biosynthetic and acquisition (THI) genes during starvation. There have not been genome-wide analyses of Pdc2 binding. Previously, we identified small regions of Pdc2-regulated genes sufficient to confer thiamine regulation. Here, we performed deletion analyses on these regions. We observed that when the S. cerevisiae PDC5 promoter is introduced into N. glabratus, it is thiamine starvation inducible but does not require the Thi3 coregulator. The ScPDC5 promoter contains a 22-bp duplication with an AT-rich spacer between the 2 repeats, which are important for regulation. Loss of the first 22-bp element does not eliminate regulation, but the promoter becomes Thi3 dependent, suggesting cis architecture can generate a Thi3-independent, thiamine starvation inducible response. Whereas many THI promoters only have 1 copy of this element, addition of the first 22-bp element to a Thi3-dependent promoter confers Thi3 independence. Finally, we performed fluorescence anisotropy and chromatin immunoprecipitation sequencing. Pdc2 and Thi3 bind to regions that share similarity to the 22-bp element in the ScPDC5 promoter and previously identified cis elements in N. glabratus promoters. Also, while Pdc2 binds to THI and PDC promoters, neither Pdc2 nor Thi3 appears to bind the evolutionarily new NgPMU3 promoter that is regulated by Pdc2. Further study is warranted because PMU3 is required for cells to acquire thiamine from environments where thiamine is phosphorylated, such as in the human bloodstream.

show abstract

A NovelcisElement Achieves the Same Solution as an AncestralcisElement During Thiamine Starvation inCandida glabrata

Cited by 3 publications

References 40 publications

Pyruvate decarboxylase and thiamine biosynthetic genes are regulated differently by Pdc2 in S. cerevisiae and C. glabrata

Pyruvate decarboxylase and thiamine biosynthetic genes are regulated differently by Pdc2 in S. cerevisiae and C. glabrata

The cloned SPI-1 type 3 secretion system can be functionally expressed outside Salmonella backgrounds

Regulation of thiamine and pyruvate decarboxylase genes by Pdc2 in Nakaseomyces glabratus (Candida glabrata) is complex

Contact Info

Product

Resources

About