Ribosylurea Accumulates in Yeast<i>urc4</i>Mutants

Björnberg, Olof; Vodnala, Munender; Domkin, V. D.; Hofer, Anders; Rasmussen, Anna Andersson; Andersen, Gorm; Piškur, Jure

doi:10.1080/15257771003741265

Cited by 11 publications

(14 citation statements)

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“…Interestingly, also among the URC loci are two unknown genes, namely URC1 and URC4. Our hypothesis is that URC1 is responsible for the opening of the pyrimidine ring and the substrate is likely ribosylated, and the products are ribosyl-urea [4] and a 3-carbon product. The ribosyl-urea would then be degraded through the action of Urc4p to urea and ribose.…”

Section: Introductionmentioning

confidence: 99%

A New Expression Vector for Production of Enzymes in the YeastSaccharomyces (Lachancea) Kluyveri

Rasmussen

Schnackerz

et al. 2011

Nucleosides, Nucleotides and Nucleic Acids

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We overexpressed and purified enzymes involved in the pyrimidine catabolic pathway in the yeast Saccharomyces (Lachancea) kluyveri. A new vector was therefore designed, providing the first specific expression system in Saccharomyces kluyveri. The URC1 gene was overexpressed and a soluble protein obtained and successfully purified using the C-terminally added His-tag. Our system will be used for further studies of the structure and function of the enzymes belonging to the URC pyrimidine degradation pathway.

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Section: Introductionmentioning

confidence: 99%

A New Expression Vector for Production of Enzymes in the YeastSaccharomyces (Lachancea) Kluyveri

Rasmussen

Schnackerz

et al. 2011

Nucleosides, Nucleotides and Nucleic Acids

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“…[26] We know that formaldehyde and Ca(OH) 2 can give sugars by the formose reaction, [27][28][29] as first described by Butlerov. [4,5] As urea itself has even been shown to react directly with ribose under mildly acidic conditions, [34] we asked whether the construction of informational base-pairing systems based on the pyrolysis products of urea, biuret and triuret (Figure 1b), [35] would be possible. [31][32][33] We also know that urea was one of the most likely nitrogen-containing molecules present on the early Earth and that it can activate mineral phosphate to achieve phosphorylations.…”

Section: Introductionmentioning

confidence: 99%

Proto‐Urea‐RNA (Wöhler RNA) Containing Unusually Stable Urea Nucleosides

et al. 2019

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The RNAw orld hypothesis assumes that life on Earth began with nucleotides that formed information-carrying RNAo ligomers able to self-replicate.P rebiotic reactions leading to the contemporary nucleosides are now known, but their execution often requires specific starting materials and lengthy reaction sequences.I tw as therefore proposed that the RNAw orld was likely proceeded by ap roto-RNAw orld constructed from molecules that were likely present on the early Earth in greater abundance.H erein, we show that the prebiotic starting molecules bis-urea (biuret) and tris-urea (triuret) are able to directly react with ribose.T he urearibosides are remarkably stable because they are held together by an etwork of intramolecular,b ifurcated hydrogen bonds. This even allowed the synthesis of phosphoramidite building blocks and incorporation of the units into RNA. Investigations of the nucleotides base-pairing potential showed that triuret:G RNAb ase pairs closely resemble U:Gw obble base pairs. Based on the probable abundance of urea on the early Earth, we postulate that urea-containing RNAb ases are good candidates for aproto-RNAworld. Figure 1. a) The chemical structures of the canonical RNA nucleosides. b) The chemical structures of biuret and triuret. c) Depiction of the urea-based nucleosidesw ith potentially stabilizing hydrogen bonds.

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“…For cDNA synthesis, SuperScript III reverse transcriptase (RT; Invitrogen) was used, with 5 g of total RNA as the template and the oligo(dT) [12][13][14][15][16][17][18] primers (Invitrogen). The cDNA was labeled using a Pronto!…”

Section: Methodsmentioning

confidence: 99%

“…Uracil is transformed by the action of the URC1 and URC6 gene products and possibly one or several other enzymes. This results in the formation of 3-hydroxypropionate, which is excreted, and (phospho)ribosyl urea, which is somehow degraded to urea (17,18). Urc4p has been proposed to catalyze the degradation of the (phospho)ribosyl urea.…”

Section: Figmentioning

confidence: 99%

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Global Expression Analysis of the Yeast Lachancea (Saccharomyces) kluyveri Reveals New URC Genes Involved in Pyrimidine Catabolism

et al. 2014

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bPyrimidines are important nucleic acid precursors which are constantly synthesized, degraded, and rebuilt in the cell. Four degradation pathways, two of which are found in eukaryotes, have been described. One of them, the URC pathway, has been initially discovered in our laboratory in the yeast Lachancea kluyveri. Here, we present the global changes in gene expression in L. kluyveri in response to different nitrogen sources, including uracil, uridine, dihydrouracil, and ammonia. The expression pattern of the known URC genes, URC1-6, helped to identify nine putative novel URC genes with a similar expression pattern. The microarray analysis provided evidence that both the URC and PYD genes are under nitrogen catabolite repression in L. kluyveri and are induced by uracil or dihydrouracil, respectively. We determined the function of URC8, which was found to catalyze the reduction of malonate semialdehyde to 3-hydroxypropionate, the final degradation product of the pathway. The other eight genes studied were all putative permeases. Our analysis of double deletion strains showed that the L. kluyveri Fui1p protein transported uridine, just like its homolog in Saccharomyces cerevisiae, but we demonstrated that is was not the only uridine transporter in L. kluyveri. We also showed that the L. kluyveri homologs of DUR3 and FUR4 do not have the same function that they have in S. cerevisiae, where they transport urea and uracil, respectively. In L. kluyveri, both of these deletion strains grew normally on uracil and urea.

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Ribosylurea Accumulates in Yeasturc4Mutants

Cited by 11 publications

References 4 publications

A New Expression Vector for Production of Enzymes in the YeastSaccharomyces (Lachancea) Kluyveri

A New Expression Vector for Production of Enzymes in the YeastSaccharomyces (Lachancea) Kluyveri

Proto‐Urea‐RNA (Wöhler RNA) Containing Unusually Stable Urea Nucleosides

Global Expression Analysis of the Yeast Lachancea (Saccharomyces) kluyveri Reveals New URC Genes Involved in Pyrimidine Catabolism

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