1887

Abstract

Phosphoribosyl pyrophosphate synthetase, which is encoded by the Prs gene, catalyses the reaction of ribose-5-phosphate and adenine ribonucleotide triphosphate (ATP) and has central importance in cellular metabolism. However, knowledge about how Prs family members function and contribute to total 5-phosphoribosyl-α-1-pyrophosphate (PRPP) synthetase activity is limited. In this study, we identified that the filamentous fungus genome contains three PRPP synthase-homologous genes (, and ), among which and but not are auxotrophic genes. Transcriptional expression profiles revealed that the mRNA levels of , and are dynamic during germination, hyphal growth and sporulation and that they all showed abundant expression during the vigorous hyphal growth time point. Inhibiting the expression of or in conditional strains produced more effects on the total PRPP synthetase activity than did inhibiting , thus indicating that different AnPrs proteins are unequal in their contributions to Prs enzyme activity. In addition, the constitutive overexpression of or could significantly rescue the defective phenotype of the -absent strain, suggesting that the function of is not a specific consequence of this auxotrophic gene but instead comes from the contribution of Prs proteins to PRPP synthetase activity.

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2017-02-01
2019-12-11
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References

  1. Khorana HG, Fernandes JF, Kornberg A. Pyrophosphorylation of ribose 5-phosphate in the enzymatic synthesis of 5-phosphorylribose 1-pyrophosphate. J Biol Chem 1958;230:941–948[PubMed]
    [Google Scholar]
  2. Hove-Jensen B. Mutation in the phosphoribosylpyrophosphate synthetase gene (prs) that results in simultaneous requirements for purine and pyrimidine nucleosides, nicotinamide nucleotide, histidine, and tryptophan in Escherichia coli. J Bacteriol 1988;170:1148–1152 [CrossRef][PubMed]
    [Google Scholar]
  3. Krath BN, Eriksen TA, Poulsen TS, Hove-Jensen B. Cloning and sequencing of cDNAs specifying a novel class of phosphoribosyl diphosphate synthase in Arabidopsis thaliana. Biochim Biophys Acta 1999;1430:403–408 [CrossRef][PubMed]
    [Google Scholar]
  4. Jiménez A, Santos MA, Revuelta JL. Phosphoribosyl pyrophosphate synthetase activity affects growth and riboflavin production in Ashbya gossypii. BMC Biotechnol 2008;8:67 [CrossRef][PubMed]
    [Google Scholar]
  5. Fang H, Xie X, Xu Q, Zhang C, Chen N. Enhancement of cytidine production by coexpression of gnd, zwf, and prs genes in recombinant Escherichia coli CYT15. Biotechnol Lett 2013;35:245–251 [CrossRef][PubMed]
    [Google Scholar]
  6. Breda A, Martinelli LK, Bizarro CV, Rosado LA, Borges CB et al. Wild-type phosphoribosylpyrophosphate synthase (PRS) from Mycobacterium tuberculosis: a bacterial class II PRS?. PLoS One 2012;7:e39245 [CrossRef][PubMed]
    [Google Scholar]
  7. Hove-Jensen B. Phosphoribosylpyrophosphate (PRPP)-less mutants of Escherichia coli. Mol Microbiol 1989;3:1487–1492 [CrossRef][PubMed]
    [Google Scholar]
  8. Shi S, Chen T, Zhang Z, Chen X, Zhao X. Transcriptome analysis guided metabolic engineering of Bacillus subtilis for riboflavin production. Metab Eng 2009;11:243–252 [CrossRef][PubMed]
    [Google Scholar]
  9. Becker MA, Taylor W, Smith PR, Ahmed M. Overexpression of the normal phosphoribosylpyrophosphate synthetase 1 isoform underlies catalytic superactivity of human phosphoribosylpyrophosphate synthetase. J Biol Chem 1996;271:19894–19899 [CrossRef][PubMed]
    [Google Scholar]
  10. de Brouwer AP, van Bokhoven H, Nabuurs SB, Arts WF, Christodoulou J et al. PRPS1 mutations: four distinct syndromes and potential treatment. Am J Hum Genet 2010;86:506–518 [CrossRef][PubMed]
    [Google Scholar]
  11. Kim HJ, Sohn KM, Shy ME, Krajewski KM, Hwang M et al. Mutations in PRPS1, which encodes the phosphoribosyl pyrophosphate synthetase enzyme critical for nucleotide biosynthesis, cause hereditary peripheral neuropathy with hearing loss and optic neuropathy (CMTX5). Am J Hum Genet 2007;81:552–558 [CrossRef][PubMed]
    [Google Scholar]
  12. Roessler BJ, Nosal JM, Smith PR, Heidler SA, Palella TD et al. Human X-linked phosphoribosylpyrophosphate synthetase superactivity is associated with distinct point mutations in the PRPS1 gene. J Biol Chem 1993;268:26476–26481[PubMed]
    [Google Scholar]
  13. Yen RC, Adams WB, Lazar C, Becker MA. Evidence for X-linkage of human phosphoribosylpyrophosphate synthetase. Proc Natl Acad Sci USA 1978;75:482–485 [CrossRef][PubMed]
    [Google Scholar]
  14. Li S, Lu Y, Peng B, Ding J. Crystal structure of human phosphoribosylpyrophosphate synthetase 1 reveals a novel allosteric site. Biochem J 2007;401:39–47 [CrossRef][PubMed]
    [Google Scholar]
  15. Liu X, Han D, Li J, Han B, Ouyang X et al. Loss-of-function mutations in the PRPS1 gene cause a type of nonsyndromic X-linked sensorineural deafness, DFN2. Am J Hum Genet 2010;86:65–71 [CrossRef][PubMed]
    [Google Scholar]
  16. Mittal R, Patel K, Mittal J, Chan B, Yan D et al. Association of PRPS1 mutations with disease phenotypes. Dis Markers 2015;2015:1–7 [CrossRef]
    [Google Scholar]
  17. Alderwick LJ, Lloyd GS, Ghadbane H, May JW, Bhatt A et al. The C-terminal domain of the Arabinosyltransferase Mycobacterium tuberculosis EmbC is a lectin-like carbohydrate binding module. PLoS Pathog 2011;7:e1001299 [CrossRef][PubMed]
    [Google Scholar]
  18. Lucarelli AP, Buroni S, Pasca MR, Rizzi M, Cavagnino A et al. Mycobacterium tuberculosis phosphoribosylpyrophosphate synthetase: biochemical features of a crucial enzyme for mycobacterial cell wall biosynthesis. PLoS One 2010;5:e15494 [CrossRef][PubMed]
    [Google Scholar]
  19. Krath BN, Hove-Jensen B. Organellar and cytosolic localization of four phosphoribosyl diphosphate synthase isozymes in Spinach. Plant Physiol 1999;119:497–506 [CrossRef][PubMed]
    [Google Scholar]
  20. Ugbogu EA, Wippler S, Euston M, Kouwenhoven EN, de Brouwer AP et al. The contribution of the nonhomologous region of Prs1 to the maintenance of cell wall integrity and cell viability. FEMS Yeast Res 2013;13:291–301 [CrossRef][PubMed]
    [Google Scholar]
  21. Mateos L, Jiménez A, Revuelta JL, Santos MA. Purine biosynthesis, riboflavin production, and trophic-phase span are controlled by a Myb-related transcription factor in the fungus Ashbya gossypii. Appl Environ Microbiol 2006;72:5052–5060 [CrossRef][PubMed]
    [Google Scholar]
  22. Carter AT, Beiche F, Hove-Jensen B, Narbad A, Barker PJ et al. PRS1 is a key member of the gene family encoding phosphoribosylpyrophosphate synthetase in Saccharomyces cerevisiae. Mol Gen Genet 1997;254:148–156 [CrossRef][PubMed]
    [Google Scholar]
  23. Hernando Y, Parr A, Schweizer M. PRS5, the fifth member of the phosphoribosyl pyrophosphate synthetase gene family in Saccharomyces cerevisiae, is essential for cell viability in the absence of either PRS1 or PRS3. J Bacteriol 1998;180:6404–6407[PubMed]
    [Google Scholar]
  24. Hove-Jensen B. Heterooligomeric phosphoribosyl diphosphate synthase of Saccharomyces cerevisiae: combinatorial expression of the five PRS genes in Escherichia coli. J Biol Chem 2004;279:40345–40350 [CrossRef][PubMed]
    [Google Scholar]
  25. Kleineidam A, Vavassori S, Wang K, Schweizer LM, Griac P et al. Valproic acid- and lithium-sensitivity in prs mutants of Saccharomyces cerevisiae. Biochem Soc Trans 2009;37:1115–1120 [CrossRef][PubMed]
    [Google Scholar]
  26. Schneiter R, Carter AT, Hernando Y, Zellnig G, Schweizer LM et al. The importance of the five phosphoribosyl-pyrophosphate synthetase (Prs) gene products of Saccharomyces cerevisiae in the maintenance of cell integrity and the subcellular localization of Prs1p. Microbiology 2000;146:3269–3278 [CrossRef][PubMed]
    [Google Scholar]
  27. Vavassori S, Wang K, Schweizer LM, Schweizer M. In Saccharomyces cerevisiae, impaired PRPP synthesis is accompanied by valproate and Li+ sensitivity. Biochem Soc Trans 2005;33:1154–1157 [CrossRef][PubMed]
    [Google Scholar]
  28. Wang K, Vavassori S, Schweizer LM, Schweizer M. Impaired PRPP-synthesizing capacity compromises cell integrity signalling in Saccharomyces cerevisiae. Microbiology 2004;150:3327–3339 [CrossRef][PubMed]
    [Google Scholar]
  29. Zhong G, Wei W, Guan Q, Ma Z, Wei H et al. Phosphoribosyl pyrophosphate synthetase, as a suppressor of the sepH mutation in Aspergillus nidulans, is required for the proper timing of septation. Mol Microbiol 2012;86:894–907 [CrossRef][PubMed]
    [Google Scholar]
  30. Gupta SK, Maggon KK, Venkitasubramanian TA. Effect of zinc on adenine nucleotide pools in relation to aflatoxin biosynthesis in Aspergillus parasiticus. Appl Environ Microbiol 1976;32:753–756[PubMed]
    [Google Scholar]
  31. Käfer E. Meiotic and mitotic recombination in Aspergillus and its chromosomal aberrations. Adv Genet 1977;19:33–131[PubMed]
    [Google Scholar]
  32. Moreno S, Klar A, Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol 1991;194:795–823[PubMed][CrossRef]
    [Google Scholar]
  33. Wang J, Hu H, Wang S, Shi J, Chen S et al. The important role of actinin-like protein (AcnA) in cytokinesis and apical dominance of hyphal cells in Aspergillus nidulans. Microbiology 2009;155:2714–2725 [CrossRef][PubMed]
    [Google Scholar]
  34. Wang PM, Choera T, Wiemann P, Pisithkul T, Amador-Noguez D et al. TrpE feedback mutants reveal roadblocks and conduits toward increasing secondary metabolism in Aspergillus fumigatus. Fungal Genet Biol 2016;89:102–113 [CrossRef][PubMed]
    [Google Scholar]
  35. Zhong GW, Jiang P, Qiao WR, Zhang YW, Wei WF et al. Protein phosphatase 2a (PP2A) regulatory subunits ParA and PabA orchestrate septation and conidiation and are essential for PP2A activity in Aspergillus nidulans. Eukaryot Cell 2014;13:1494–1506 [CrossRef][PubMed]
    [Google Scholar]
  36. Wang G, Lu L, Zhang CY, Singapuri A, Yuan S. Calmodulin concentrates at the apex of growing hyphae and localizes to the Spitzenkörper in Aspergillus nidulans. Protoplasma 2006;228:159–166 [CrossRef][PubMed]
    [Google Scholar]
  37. May GS. The highly divergent beta-tubulins of Aspergillus nidulans are functionally interchangeable. J Cell Biol 1989;109:2267–2274 [CrossRef][PubMed]
    [Google Scholar]
  38. Osmani SA, Pu RT, Morris NR. Mitotic induction and maintenance by overexpression of a G2-specific gene that encodes a potential protein kinase. Cell 1988;53:237–244 [CrossRef][PubMed]
    [Google Scholar]
  39. Yu JH, Hamari Z, Han KH, Seo JA, Reyes-Domínguez Y et al. Double-joint PCR: a PCR-based molecular tool for gene manipulations in filamentous fungi. Fungal Genet Biol 2004;41:973–981 [CrossRef][PubMed]
    [Google Scholar]
  40. Liu B, Xiang X, Lee YR. The requirement of the LC8 dynein light chain for nuclear migration and septum positioning is temperature dependent in Aspergillus nidulans. Mol Microbiol 2003;47:291–301 [CrossRef][PubMed]
    [Google Scholar]
  41. Osmani AH, Oakley BR, Osmani SA. Identification and analysis of essential Aspergillus nidulans genes using the heterokaryon rescue technique. Nat Protoc 2006;1:2517–2526 [CrossRef][PubMed]
    [Google Scholar]
  42. Harris SD, Morrell JL, Hamer JE. Identification and characterization of Aspergillus nidulans mutants defective in cytokinesis. Genetics 1994;136:517–532[PubMed]
    [Google Scholar]
  43. Shi J, Chen W, Liu Q, Chen S, Hu H et al. Depletion of the MobB and CotA complex in Aspergillus nidulans causes defects in polarity maintenance that can be suppressed by the environment stress. Fungal Genet Biol 2008;45:1570–1581 [CrossRef][PubMed]
    [Google Scholar]
  44. Zhang Y, Zheng Q, Sun C, Song J, Gao L et al. Palmitoylation of the cysteine residue in the DHHC motif of a palmitoyl transferase mediates Ca2+ homeostasis in Aspergillus. PLoS Genet 2016;12:e1005977 [CrossRef][PubMed]
    [Google Scholar]
  45. Cai ZD, Chai YF, Zhang CY, Qiao WR, Sang H et al. The Gβ-like protein CpcB is required for hyphal growth, conidiophore morphology and pathogenicity in Aspergillus fumigatus. Fungal Genet Biol 2015;81:120–131 [CrossRef][PubMed]
    [Google Scholar]
  46. Liu FF, Pu L, Zheng QQ, Zhang YW, Gao RS et al. Calcium signaling mediates antifungal activity of triazole drugs in the Aspergilli. Fungal Genet Biol 2015;81:182–190 [CrossRef][PubMed]
    [Google Scholar]
  47. Koenigsknecht MJ, Fenlon LA, Downs DM. Phosphoribosylpyrophosphate synthetase (PrsA) variants alter cellular pools of ribose 5-phosphate and influence thiamine synthesis in Salmonella enterica. Microbiology 2010;156:950–959 [CrossRef][PubMed]
    [Google Scholar]
  48. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–254 [CrossRef][PubMed]
    [Google Scholar]
  49. Song J, Zhai P, Zhang Y, Zhang C, Sang H et al. The Aspergillus fumigatus damage resistance protein family coordinately regulates ergosterol biosynthesis and azole susceptibility. MBio 2016;7:e01919-15 [CrossRef][PubMed]
    [Google Scholar]
  50. Breakspear A, Momany M. Aspergillus nidulans conidiation genes dewA, fluG, and stuA are differentially regulated in early vegetative growth. Eukaryot Cell 2007;6:1697–1700 [CrossRef][PubMed]
    [Google Scholar]
  51. Suh MJ, Fedorova ND, Cagas SE, Hastings S, Fleischmann RD et al. Development stage-specific proteomic profiling uncovers small, lineage specific proteins most abundant in the Aspergillus Fumigatus conidial proteome. Proteome Sci 2012;10:30 [CrossRef][PubMed]
    [Google Scholar]
  52. Chen P, Gao R, Chen S, Pu L, Li P et al. A pericentrin-related protein homolog in Aspergillus nidulans plays important roles in nucleus positioning and cell polarity by affecting microtubule organization. Eukaryot Cell 2012;11:1520–1530 [CrossRef][PubMed]
    [Google Scholar]
  53. Hernando Y, Carter AT, Parr A, Hove-Jensen B, Schweizer M. Genetic analysis and enzyme activity suggest the existence of more than one minimal functional unit capable of synthesizing phosphoribosyl pyrophosphate in Saccharomyces cerevisiae. J Biol Chem 1999;274:12480–12487 [CrossRef][PubMed]
    [Google Scholar]
  54. Ugbogu EA, Wang K, Schweizer LM, Schweizer M. Metabolic gene products have evolved to interact with the cell wall integrity pathway in Saccharomyces cerevisiae. FEMS Yeast Res 2016;16:fow092 [CrossRef][PubMed]
    [Google Scholar]
  55. Nayak T, Szewczyk E, Oakley CE, Osmani A, Ukil L et al. A versatile and efficient gene-targeting system for Aspergillus nidulans. Genetics 2006;172:1557–1566 [CrossRef][PubMed]
    [Google Scholar]
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