Molecular cloning and characterization of theCandida albicansUBI3 gene coding for a ubiquitin-hybrid protein

Roig, Patricia; Martinez, Josè; Gil, M. Luisa; Gozalbo, Daniel

doi:10.1002/1097-0061(200011)16:15<1413::aid-yea632>3.0.co;2-u

Cited by 16 publications

(32 citation statements)

References 29 publications

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“…The polyubiquitin gene UBI4 is induced in response to numerous stress responses, including heat, peroxide, and cell wall stresses (88). This contrasts with a ubiquitin hybrid protein, Ubi3, which is involved in ribosome biogenesis and which is coordinately regulated with components of the translational apparatus (145,147).…”

Section: Reversible Peptide Modificationsmentioning

confidence: 99%

Posttranslational Modifications of Proteins in the Pathobiology of Medically Relevant Fungi

Leach

Brown

2012

Eukaryot Cell

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Posttranslational modifications of proteins drive a wide variety of cellular processes in eukaryotes, regulating cell growth and division as well as adaptive and developmental processes. With regard to the fungal kingdom, most information about posttranslational modifications has been generated through studies of the model yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe, where, for example, the roles of protein phosphorylation, glycosylation, acetylation, ubiquitination, sumoylation, and neddylation have been dissected. More recently, information has begun to emerge for the medically important fungal pathogens Candida albicans, Aspergillus fumigatus, and Cryptococcus neoformans, highlighting the relevance of posttranslational modifications for virulence. We review the available literature on protein modifications in fungal pathogens, focusing in particular upon the reversible peptide modifications sumoylation, ubiquitination, and neddylation.

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Section: Reversible Peptide Modificationsmentioning

confidence: 99%

Posttranslational Modifications of Proteins in the Pathobiology of Medically Relevant Fungi

Leach

Brown

2012

Eukaryot Cell

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Section: Introductionmentioning

confidence: 99%

“…UBI3 encodes a ubiquitinhybrid protein involved in ribosome biogenesis, and UBI4 encodes polyubiquitin [19,20,22]. Different mechanisms regulate the expression of these genes; UBI3 is expressed coordinately with members of the translational apparatus according to the requirements of protein synthesis and cellular growth rate, whereas UBI4 expression is enhanced by stress conditions [19]. Regulatory mechanisms such as the control of mRNA stability may be involved in the regulation of gene expression, especially in the case of UBI3 and ribosomal protein genes in which a stress (heat)-induced mRNAdecay mechanism may participate in their regulation [13,17,19].…”

Section: Introductionmentioning

confidence: 99%

Candida albicans UBI3 and UBI4 promoter regions confer differential regulation of invertase production to Saccharomyces cerevisiae cells in response to stress

Roig

Gozalbo

2002

Int Microbiol

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Candida albicans ubiquitin genes UBI3 and UBI4 encode a ubiquitin-hybrid protein involved in ribosome biogenesis and polyubiquitin, respectively. In this work we show that UBI3 and UBI4 promoter regions confer differential expression consistent with the function of their encoded gene products. Hybrid genes were constructed containing the SUC2 coding region under the control of UBI3 or UBI4 promoters in the yeast vector pLC7. Invertase production in Saccharomyces cerevisiae transformants was differentially regulated: the UBI4 promoter was induced by stress conditions (thermal upshift and/or starvation) whereas the UBI3 promoter conferred constitutive invertase production in growing yeast cells. These results indicate that the UBI4 promoter is regulated by stress-response signaling pathways, whereas the UBI3 promoter is controlled according to the requirement for protein synthesis to support cell growth.

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“…cerevisiae SEY 2101 was transformed with pTDH, pPRO, pINV, pDEL1-7 or pSC1, basically as previously described (Maneu et al, 2000;Roig et al, 2000). Transformants were isolated on selective media lacking uracil.…”

Section: Yeast Transformationmentioning

confidence: 99%

“…Total RNA from S. cerevisiae SEY 2101 and a selected transformant containing pTDH, was isolated from exponentially growing cells cultured in YPD at 28 • C, as previously described (Maneu et al, 1996;Roig et al, 2000). Agarose gel electrophoresis of RNA samples and transfer to Nylon filters were performed according to standard procedures (Sambrook et al, 1989), and for hybridization and chemiluminescent detection the Lumigen PPD kit (Roche Diagnostics, Mannheim) was used.…”

Section: Northern Blotmentioning

confidence: 99%

Candida albicans TDH3 gene promotes secretion of internal invertase when expressed in Saccharomyces cerevisiae as a glyceraldehyde‐3‐phosphate dehydrogenase‐invertase fusion protein

Delgado

Gil

Gozalbo

2003

Yeast

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We have checked the ability of the Candida albicans GAPDH polypeptide, which lacks a conventional N-terminal signal peptide, to reach the cell wall in Saccharomyces cerevisiae by using an intracellular form of the yeast invertase as a reporter protein.A hybrid TDH3-SUC2 gene containing the C. albicans TDH3 promoter sequences and a coding region encoding a fusion protein formed by the C. albicans GAPDH polypeptide, fused at its C-terminus with the yeast internal invertase, was constructed in a centromer derivative plasmid and transformed into a Suc − S. cerevisiae strain. Transformants displayed invertase activity measured in intact whole cells, and were able to grow on sucrose as the sole fermentable carbon source. Northern blot analysis with both TDH3 and SUC2 probes detected a single mRNA species of the expected size (about 2.7 kb), and Western immunoblot analysis of cell-free extracts, using a monoclonal antibody (mAb49) against a C. albicans GAPDH epitope, showed the presence of a 90 kDa polypeptide corresponding to the GAPDH-invertase fusion protein. This indicates that the TDH3 gene is able to direct part of the encoded gene product to the cell wall, and that any putative motifs for this targeting should be within the GAPDH amino acid sequence. Further analysis, using the same approach, of a panel of seven N-and C-terminal GAPDH truncates revealed that the region required for the cell wall targeting is located within the N-terminal half of the protein.

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Molecular cloning and characterization of theCandida albicansUBI3 gene coding for a ubiquitin-hybrid protein

Cited by 16 publications

References 29 publications

Posttranslational Modifications of Proteins in the Pathobiology of Medically Relevant Fungi

Posttranslational Modifications of Proteins in the Pathobiology of Medically Relevant Fungi

Candida albicans UBI3 and UBI4 promoter regions confer differential regulation of invertase production to Saccharomyces cerevisiae cells in response to stress

Candida albicans TDH3 gene promotes secretion of internal invertase when expressed in Saccharomyces cerevisiae as a glyceraldehyde‐3‐phosphate dehydrogenase‐invertase fusion protein

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