Phosphoribosylpyrophosphate synthetase (<i>PRS</i>): A new gene family in <i>Saccharomyces cerevisiae</i>

Carter, Andrew T.; Narbad, Arjan; Pearson, Bruce M.; Beck, Karl‐Friedrich; Baum, Bobby; Logghe, Marc; Contreras, Roland; Schweizer, Michael

doi:10.1002/yea.320100805

Cited by 39 publications

(50 citation statements)

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“…The inserts of all the plasmids constructed with PCR-amplified DNA fragments were sequenced. The deduced amino acid sequences of S. cerevisiae PRS1, PRS2, PRS3, and PRS5 were identical to those published previously, whereas the nucleotide sequence of the PRS4 gene used in the present work differed slightly from that published previously (4,5). 2 2 The nucleotide sequence of the PRS4 gene used in the present work differed slightly from that published previously.…”

Section: Methodssupporting

confidence: 70%

Heterooligomeric Phosphoribosyl Diphosphate Synthase of Saccharomyces cerevisiae

Hove‐Jensen

2004

Journal of Biological Chemistry

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The yeast Saccharomyces cerevisiae contains five phosphoribosyl diphosphate (PRPP) synthase-homologous genes (PRS1-5), which specify PRPP synthase subunits 1-5. Expression of the five S. cerevisiae PRS genes individually in an Escherichia coli PRPP-less strain (⌬prs) showed that a single PRS gene product had no PRPP synthase activity. In contrast, expression of five pairwise combinations of PRS genes resulted in the formation of active PRPP synthase. These combinations were PRS1 PRS2, PRS1 PRS3, and PRS1 PRS4, as well as PRS5 PRS2 and PRS5 PRS4. None of the remaining five possible pairwise combinations of PRS genes appeared to produce active enzyme. Extract of an E. coli strain containing a plasmid-borne PRS1 gene and a chromosome-borne PRS3 gene contained detectable PRPP synthase activity, whereas extracts of strains containing PRS1 PRS2, PRS1 PRS4, PRS5 PRS2, or PRS5 PRS4 contained no detectable PRPP synthase activity. In contrast PRPP could be detected in growing cells containing PRS1 PRS2, PRS1 PRS3, PRS5 PRS2, or PRS5 PRS4. These apparent conflicting results indicate that, apart from the PRS1 PRS3-specified enzyme, PRS-specified enzyme is functional in vivo but unstable when released from the cell. Certain combinations of three PRS genes appeared to produce an enzyme that is stable in vitro. Thus, extracts of strains harboring PRS1 PRS2 PRS5, PRS1 PRS4 PRS5, or PRS2 PRS4 PRS5 as well as extracts of strains harboring combinations with PRS1 PRS3 contained readily assayable PRPP synthase activity. The data indicate that although certain pairwise combinations of subunits produce an active enzyme, yeast PRPP synthase requires at least three different subunits to be stable in vitro. The activity of PRPP synthases containing subunits 1 and 3 or subunits 1, 2, and 5 was found to be dependent on P i , to be temperature-sensitive, and inhibited by ADP.is an important component of the metabolism of most organisms. PRPP is a precursor of the biosynthesis of purine, pyrimidine, and pyridine nucleotides, as well as of the amino acids tryptophan and histidine (1). The biosynthesis of PRPP is catalyzed by PRPP synthase (ATP: D-ribose 5-phosphate pyrophosphotransferase, EC 2.7.6.1), which is encoded by a PRS gene: ribose 5-phosphate ϩ ATP 3 PRPP ϩ AMP (2). Except for certain specialized mutants of Escherichia coli, all freeliving organisms contain at least one gene encoding PRPP synthase. These E. coli mutants require guanosine, uridine, histidine, tryptophan, and NAD (1, 3). The yeast Saccharomyces cerevisiae contains five PRPP synthase-homologous genes, PRS1-5, encoding PRPP synthase subunits 1-5, respectively. All five genes have been shown to be expressed (4 -6). Phylogenetic analysis revealed a close relationship of S. cerevisiae PRPP synthase subunits with PRPP synthases from other eukaryotic organisms, including human, rat, and Caenorhabditis elegans. S. cerevisiae PRPP synthase subunits 2, 3, and 4 resemble the "classical" class I PRPP synthases from E. coli, Bacillus subtilis, and human in length (311-355 amino aci...

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Section: Methodssupporting

confidence: 70%

Heterooligomeric Phosphoribosyl Diphosphate Synthase of Saccharomyces cerevisiae

Hove‐Jensen

2004

Journal of Biological Chemistry

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“…A systematic phenotypic analysis has been carried out with our collection of strains representing all possible combinations of deletions of the five PRS genes. The results obtained define three phenotypes: (i) a synthetic lethal phenotype when PRS1 or PRS3 was deleted from a prs5D strain -simultaneous deletion of PRS2 and PRS4 in combination with loss of PRS1 or PRS3 also results in inviability; (ii) a second phenotype that is encountered in strains containing deletions of PRS1 and PRS3 together or in combination with lack of PRS2 or PRS4 manifests itself as a reduction in growth rate, enzyme activity and nucleotide content; and (iii) deletion of PRS2, PRS4 or PRS5 or combinations thereof has reduced enzyme activity, but are unimpaired in growth and nucleotide profiles (Carter et al, 1994;Hernando et al, 1998Hernando et al, , 1999. Three viable triple deletion combinations, prs2D prs4D prs5D, prs1D prs3D prs4D and prs1D prs2D prs3D, define three minimal subunits, Prs1/Prs3, Prs2/Prs5, Prs4/Prs5, respectively An extensive two-hybrid (Y2H) analysis suggested the existence in the wild-type of two interacting functional entities which may have compensatory function since in the absence of one entity or one of its components the yeast cell can still survive (Hernando et al, 1998(Hernando et al, , 1999.…”

Section: Introductionmentioning

confidence: 99%

Impaired PRPP-synthesizing capacity compromises cell integrity signalling in Saccharomyces cerevisiae

et al. 2004

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“…Not surprisingly, the expression pattern of DjPRPS in the head and tail segments was same as that in the intact planarian. (a) (b) (c) [39], and five members are included in PRPS gene family of Saccharomyces cerevisiae [18,20]. All these genes have been cloned and sequenced.…”

Section: The Spatial Expression Pattern Of Djprps In Sexual Intact Anmentioning

confidence: 99%

“…As one of the comparatively conservative genes during evolution, PRPS are found ubiquitously from bacteria to human. It has been reported that PRPS genes have been cloned and sequenced from a variety of organisms, including bacteria [19], yeast [20], protozoa [21], nematodes [22], rat [24] and human [25]. However, except for Hase and co-workers [27], this important gene has not been explored in planarians so far.…”

Section: The Spatial Expression Pattern Of Djprps In Sexual Intact Anmentioning

confidence: 99%

“…PRPS is required for both the de novo and salvage pathways and therefore represents a key enzyme in intermediary metabolism [18]. As one of the more conserved genes during evolution, PRPS gene has been cloned and sequenced from a variety of organisms, including bacteria [19], yeast [20], protozoa [21], nematodes [22], plants [23], rat [24] and human [25] etc. In many organisms, PRPS can be encoded by more than one genes [26,18].…”

Section: Introductionmentioning

confidence: 99%

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Cloning and characterization of DjPRPS gene in freshwater planarian Dugesia japonica

et al. 2015

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Objective: To clone the phosphoribosyl pyrophosphate synthetase gene from Dugesia japonica, and to do the bioinformatics and spatial expression analysis of the gene. Methods:The phosphoribosyl pyrophosphate synthetase gene (DjPRPS) was cloned from Dugesia japonica using the rapid amplification of cDNA ends method and its sequence was analyzed with biological softwares. Spatial expression pattern of the gene was detected by whole-mount in situ hybridization. Results:The full length cDNA of DjPRPS was 1,086 bp, containing a 80 bp of 5'-untranslated region, a 55 bp of 3'-untranslated region and a 951 bp of open reading frame encoding a protein of 316 amino acids with a calculated molecular mass of 35.18 kDa and isoelectric point of 8.14. The deduced amino acid sequence shared the conserved eukaryotic PRPS family motif. Phylogenetic analysis supported the traditional concept of zoological taxonomy in large scale. Intense positive signals were detected as bilateral lines of patches in the ventral side where the ovaries and yolk glands located in the intact sexual planarians. In the regenerating planarians, the expression pattern was similar to that of the intact samples, and no intense transcipt signals were detected in the blastema. Conclusion: DjPRPS sequence was highly homologous with that of other eukaryotic phosphoribosyl pyrophosphate synthetase gene. Intense transcript signals of DjPRPS were not detected in the blastema but in ovaries and yolk glands, suggesting that its function was probably reproduction-related and maybe not involved in regeneration. Key Words: Dugesia japonica, DjPRPS, cloning, whole-mount in situ hybridization Conflict of Interest: The authors have no conflict of interest. ÖZETAmaç: Dugesia japonica'dan fosforibozil pirofosfat sentetaz geninin klonlanması ve genin mekansal ekspresyon analizi ile bioinformatik çalışmaların yapılmasıdır. Metod: Fosforibozil pirofosfat sentetaz (DjPRPS) geni Dugesia japonica'dan cDNA ucunun hızlı amplifikasyon metodu kullanılarak klonlandı ve dizilim biyolojik yazılımlar ile analiz edildi. Genin mekansal ekspresyon motifi whole-mount in situ hibridizasyon ile saptandı. Bulgular: DjPRPS geninin tüm cDNA uzunluğu 1,086 bp'dir. Bu yapı, 80 bp 5'-çevrilmemiş bölge, 55 bp 3'-çevrilmemiş bölge ve 951 bp'lik izoelektrik noktası 8.14 ve hesaplanan moleküler kütlesi 35.18 kDa olan 316 amino asitden oluşan proteini kodlayan açık okuma ucu içe-rir. Ortaya çıkarılan amino asit dizilimi korunmuş ökaryotik PRPS ailesi motifini paylaşmıştır. Filogenetik analizler büyük oranda geleneksel zoolojik taksonomi kavramlarını desteklemiştir. Bozulmamış seksüel planaryalarda yerleşmiş yumurtalık ve yolk keselerinde ventral bölgede yoğunlaştırılmış pozitif sinyaller bilateral yama tarzında çizgiler olarak tespit edildi. Rejenere olan planaryalarda da ifade edilme kalıbı benzerdi ve embriyonik hücre gruplarında yoğun transkripsiyon sinyalleri tespit edilmedi. Sonuç: DjPRPS gen dizilimi diğer ökaryotik fosforibozil pirofosfat sentetaz geni ile yüksek oranda homoloji göstermekte...

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Phosphoribosylpyrophosphate synthetase (PRS): A new gene family in Saccharomyces cerevisiae

Cited by 39 publications

References 39 publications

Heterooligomeric Phosphoribosyl Diphosphate Synthase of Saccharomyces cerevisiae

Heterooligomeric Phosphoribosyl Diphosphate Synthase of Saccharomyces cerevisiae

Impaired PRPP-synthesizing capacity compromises cell integrity signalling in Saccharomyces cerevisiae

Cloning and characterization of DjPRPS gene in freshwater planarian Dugesia japonica

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