BMC Microbiology | |
Comprehensive annotation of secondary metabolite biosynthetic genes and gene clusters of Aspergillus nidulans, A. fumigatus, A. niger and A. oryzae | |
Gavin Sherlock2  Jennifer R Wortman1  Farrell Wymore2  Prachi Shah2  Gustavo C Cerqueira1  Martha B Arnaud2  Marek S Skrzypek2  Jonathan Binkley2  Diane O Inglis2  | |
[1] Broad Institute, 7 Cambridge Center, Cambridge, MA 02141, USA;Department of Genetics, Stanford University Medical School, Stanford, CA 94305-5120, USA | |
关键词: Sybil; Secondary metabolism; Genome annotation; Gene Ontology; Gene clusters; Aspergillus; | |
Others : 1143863 DOI : 10.1186/1471-2180-13-91 |
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received in 2013-02-15, accepted in 2013-04-15, 发布年份 2013 | |
【 摘 要 】
Background
Secondary metabolite production, a hallmark of filamentous fungi, is an expanding area of research for the Aspergilli. These compounds are potent chemicals, ranging from deadly toxins to therapeutic antibiotics to potential anti-cancer drugs. The genome sequences for multiple Aspergilli have been determined, and provide a wealth of predictive information about secondary metabolite production. Sequence analysis and gene overexpression strategies have enabled the discovery of novel secondary metabolites and the genes involved in their biosynthesis. The Aspergillus Genome Database (AspGD) provides a central repository for gene annotation and protein information for Aspergillus species. These annotations include Gene Ontology (GO) terms, phenotype data, gene names and descriptions and they are crucial for interpreting both small- and large-scale data and for aiding in the design of new experiments that further Aspergillus research.
Results
We have manually curated Biological Process GO annotations for all genes in AspGD with recorded functions in secondary metabolite production, adding new GO terms that specifically describe each secondary metabolite. We then leveraged these new annotations to predict roles in secondary metabolism for genes lacking experimental characterization. As a starting point for manually annotating Aspergillus secondary metabolite gene clusters, we used antiSMASH (antibiotics and Secondary Metabolite Analysis SHell) and SMURF (Secondary Metabolite Unknown Regions Finder) algorithms to identify potential clusters in A. nidulans, A. fumigatus, A. niger and A. oryzae, which we subsequently refined through manual curation.
Conclusions
This set of 266 manually curated secondary metabolite gene clusters will facilitate the investigation of novel Aspergillus secondary metabolites.
【 授权许可】
2013 Inglis et al.; licensee BioMed Central Ltd.
【 预 览 】
Files | Size | Format | View |
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20150330024043246.pdf | 510KB | download | |
Figure 3. | 28KB | Image | download |
Figure 2. | 35KB | Image | download |
Figure 1. | 47KB | Image | download |
【 图 表 】
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【 参考文献 】
- [1]Bhetariya PJ, Madan T, Basir SF, Varma A, Usha SP: Allergens/Antigens, toxins and polyketides of important Aspergillus species. Indian J Clin Biochem 2011, 26:104-119.
- [2]Rohlfs M, Albert M, Keller NP, Kempken F: Secondary chemicals protect mould from fungivory. Biol Lett 2007, 3:523-525.
- [3]MacCabe AP, van Liempt H, Palissa H, Unkles SE, Riach MB, Pfeifer E, von Döhren H, Kinghorn JR: Delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase from Aspergillus nidulans. Molecular characterization of the acvA gene encoding the first enzyme of the penicillin biosynthetic pathway. J Biol Chem 1991, 266:12646-12654.
- [4]MacCabe AP, Riach MB, Unkles SE, Kinghorn JR: The Aspergillus nidulans npeA locus consists of three contiguous genes required for penicillin biosynthesis. EMBO J 1990, 9:279-287.
- [5]Ramón D, Carramolino L, Patiño C, Sánchez F, Peñalva MA: Cloning and characterization of the isopenicillin N synthetase gene mediating the formation of the beta-lactam ring in Aspergillus nidulans. Gene 1987, 57:171-181.
- [6]Yu JH, Leonard TJ: Sterigmatocystin biosynthesis in Aspergillus nidulans requires a novel type I polyketide synthase. J Bacteriol 1995, 177:4792-4800.
- [7]Keller NP, Segner S, Bhatnagar D, Adams TH: stcS, a putative P-450 monooxygenase, is required for the conversion of versicolorin A to sterigmatocystin in Aspergillus nidulans. Appl Environ Microbiol 1995, 61:3628-3632.
- [8]Kelkar HS, Keller NP, Adams TH: Aspergillus nidulans stcP encodes an O-methyltransferase that is required for sterigmatocystin biosynthesis. Appl Environ Microbiol 1996, 62:4296-4298.
- [9]Butchko RA, Adams TH, Keller NP: Aspergillus nidulans mutants defective in stc gene cluster regulation. Genetics 1999, 153:715-720.
- [10]Kelkar HS, Skloss TW, Haw JF, Keller NP, Adams TH: Aspergillus nidulans stcL encodes a putative cytochrome P-450 monooxygenase required for bisfuran desaturation during aflatoxin/sterigmatocystin biosynthesis. J Biol Chem 1997, 272:1589-1594.
- [11]Luque MI, Rodríguez A, Andrade MJ, Martín A, Córdoba JJ: Development of a PCR protocol to detect aflatoxigenic molds in food products. J Food Prot 2012, 75:85-89.
- [12]Kupfahl C, Michalka A, Lass-Flörl C, Fischer G, Haase G, Ruppert T, Geginat G, Hof H: Gliotoxin production by clinical and environmental Aspergillus fumigatus strains. Int J Med Microbiol 2008, 298:319-327.
- [13]Lewis RE, Wiederhold NP, Lionakis MS, Prince RA, Kontoyiannis DP: Frequency and species distribution of gliotoxin-producing Aspergillus isolates recovered from patients at a tertiary-care cancer center. J Clin Microbiol 2005, 43:6120-6122.
- [14]Morton CO, Bouzani M, Loeffler J, Rogers TR: Direct interaction studies between Aspergillus fumigatus and human immune cells; what have we learned about pathogenicity and host immunity? Front Microbiol 2012, 3:413.
- [15]Scharf DH, Heinekamp T, Remme N, Hortschansky P, Brakhage AA, Hertweck C: Biosynthesis and function of gliotoxin in Aspergillus fumigatus. Appl Microbiol Biotechnol 2012, 93:467-472.
- [16]Andersen MR, Nielsen JB, Klitgaard A, Petersen LM, Zachariasen M, Hansen TJ, Blicher LH, Gotfredsen CH, Larsen TO, Nielsen KF, Mortensen UH: Accurate prediction of secondary metabolite gene clusters in filamentous fungi. Proc Natl Acad Sci USA 2013, 110:E99-E107.
- [17]Sanchez JF, Somoza AD, Keller NP, Wang CC: Advances in Aspergillus secondary metabolite research in the post-genomic era. Nat Prod Rep 2012, 29:351-371.
- [18]Bouhired S, Weber M, Kempf-Sontag A, Keller NP, Hoffmeister D: Accurate prediction of the Aspergillus nidulans terrequinone gene cluster boundaries using the transcriptional regulator LaeA. Fungal Genet Biol 2007, 44:1134-1145.
- [19]Perrin RM, Federova ND, Bok JW, Cramer RA, Wortman JR, Kim HS, Nierman WC, Keller NP: Transcriptional regulation of chemical diversity in Aspergillus fumigatus by LaeA. PLoS Pathog 2007, 3:523-525.
- [20]Palmer JM, Keller NP: Secondary metabolism in fungi: does chromosomal location matter? Curr Opin Microbiol 2010, 13:431-436.
- [21]Lim FY, Hou Y, Chen Y, Oh JH, Lee I, Bugni TS, Keller NP: Genome-based cluster deletion reveals an endocrocin biosynthetic pathway in Aspergillus fumigatus. Appl Environ Microbiol 2012, 78:4117-4125.
- [22]Chiang YM, Szewczyk E, Nayak T, Davidson AD, Sanchez JF, Lo HC, Ho WY, Simityan H, Kuo E, Praseuth A, Watanabe K, Oakley BR, Wang CC: Molecular genetic mining of the Aspergillus secondary metabolome: discovery of the emericellamide biosynthetic pathway. Chem Biol 2008, 15:527-532.
- [23]Ahuja M, Chiang YM, Chang SL, Praseuth MB, Entwistle R, Sanchez JF, Lo HC, Yeh HH, Oakley BR, Wang CC: Illuminating the diversity of aromatic polyketide synthases in Aspergillus nidulans. J Am Chem Soc 2012, 134:8212-8221.
- [24]Nakazawa T, Ishiuchi K, Praseuth A, Noguchi H, Hotta K, Watanabe K: Overexpressing transcriptional regulator in Aspergillus oryzae activates a silent biosynthetic pathway to produce a novel polyketide. ChemBioChem 2012, 13:855-861.
- [25]Arnaud MB, Chibucos MC, Costanzo MC, Crabtree J, Inglis DO, Lotia A, Orvis J, Shah P, Skrzypek MS, Binkley G, Miyasato SR, Wortman JR, Sherlock G: The Aspergillus Genome Database, a curated comparative genomics resource for gene, protein and sequence information for the Aspergillus research community. Nucleic Acids Res 2010, 38:D420-427.
- [26]Arnaud MB, Cerqueira GC, Inglis DO, Skrzypek MS, Binkley J, Chibucos MC, Crabtree J, Howarth C, Orvis J, Shah P, Wymore F, Binkley G, Miyasato SR, Simison M, Sherlock G, Wortman JR: The Aspergillus Genome Database (AspGD): recent developments in comprehensive multispecies curation, comparative genomics and community resources. Nucleic Acids Res 2012, 40:D653-659.
- [27]The Gene Ontology Consortium: Gene Ontology Annotations and Resources. Nucleic Acids Res 2012, 41:D530-535.
- [28]Harris MA, Clark J, Ireland A, Lomax J, Ashburner M, Foulger R, Eilbeck K, Lewis S, Marshall B, Mungall C, Richter J, Rubin GM, Blake JA, Bult C, Dolan M, Drabkin H, Eppig JT, Hill DP, Ni L, Ringwald M, Balakrishnan R, Cherry JM, Christie KR, Costanzo MC, Dwight SS, Engel S, Fisk DG, Hirschman JE, Hong EL, Nash RS, Sethuraman A, Theesfeld CL, Botstein D, Dolinski K, Feierbach B, Berardini T, Mundodi S, Rhee SY, Apweiler R, Barrell D, Camon E, Dimmer E, Lee V, Chisholm R, Gaudet P, Kibbe W, Kishore R, Schwarz EM, Sternberg P, Gwinn M, Hannick L, Wortman J, Berriman M, Wood V, de la Cruz N, Tonellato P, Jaiswal P, Seigfried T, White R, Gene Ontology Consortium: The Gene Ontology (GO) database and informatics resource. Nucleic Acids Res 2004, 32:D258-261.
- [29]Khodiyar VK, Hill DP, Howe D, Berardini TZ, Tweedie S, Talmud PJ, Breckenridge R, Bhattarcharya S, Riley P, Scambler P, Lovering RC: The representation of heart development in the gene ontology. Dev Biol 2011, 354:9-17.
- [30]Szewczyk E, Chiang YM, Oakley CE, Davidson AD, Wang CC, Oakley BR: Identification and characterization of the asperthecin gene cluster of Aspergillus nidulans. Appl Environ Microbiol 2008, 74:7607-7612.
- [31]Lo HC, Entwistle R, Guo CJ, Ahuja M, Szewczyk E, Hung JH, Chiang YM, Oakley BR, Wang CC: Two separate gene clusters encode the biosynthetic pathway for the meroterpenoids austinol and dehydroaustinol in Aspergillus nidulans. J Am Chem Soc 2012, 134:4709-4720.
- [32]Márquez-Fernández O, Trigos A, Ramos-Balderas JL, Viniegra-González G, Deising HB, Aguirre J: Phosphopantetheinyl transferase CfwA/NpgA is required for Aspergillus nidulans secondary metabolism and asexual development. Eukaryot Cell 2007, 6:710-720.
- [33]Ames BD, Haynes SW, Gao X, Evans BS, Kelleher NL, Tang Y, Walsh CT: Complexity generation in fungal peptidyl alkaloid biosynthesis: oxidation of fumiquinazoline A to the heptacyclic hemiaminal fumiquinazoline C by the flavoenzyme Af12070 from Aspergillus fumigatus. Biochemistry 2011, 50:8756-8769.
- [34]Sanchez JF, Chiang YM, Szewczyk E, Davidson AD, Ahuja M, Elizabeth Oakley C, Woo Bok J, Keller N, Oakley BR, Wang CC: Molecular genetic analysis of the orsellinic acid/F9775 gene cluster of Aspergillus nidulans. Mol Biosyst 2010, 6:587-593.
- [35]Maiya S, Grundmann A, Li X, Li SM, Turner G: Identification of a hybrid PKS/NRPS required for pseurotin A biosynthesis in the human pathogen Aspergillus fumigatus. ChemBioChem 2007, 8:1736-1743.
- [36]Sanchez JF, Entwistle R, Hung JH, Yaegashi J, Jain S, Chiang YM, Wang CC, Oakley BR: Genome-based deletion analysis reveals the prenyl xanthone biosynthesis pathway in Aspergillus nidulans. J Am Chem Soc 2011, 133:4010-4017.
- [37]Nielsen ML, Nielsen JB, Rank C, Klejnstrup ML, Holm DK, Brogaard KH, Hansen BG, Frisvad JC, Larsen TO, Mortensen UH: A genome-wide polyketide synthase deletion library uncovers novel genetic links to polyketides and meroterpenoids in Aspergillus nidulans. FEMS Microbiol Lett 2011, 321:157-166.
- [38]Khaldi N, Seifuddin FT, Turner G, Haft D, Nierman WC, Wolfe KH, Fedorova ND: SMURF: Genomic mapping of fungal secondary metabolite clusters. Fungal Genet Biol 2010, 47:736-741.
- [39]Medema MH, Blin K, Cimermancic P, de Jager V, Zakrzewski P, Fischbach MA, Weber T, Takano E, Breitling R: antiSMASH: rapid identification, annotation and analysis of secondary metabolite biosynthesis gene clusters in bacterial and fungal genome sequences. Nucleic Acids Res 2011, 39:W339-346.
- [40]Chiang YM, Szewczyk E, Davidson AD, Keller N, Oakley BR, Wang CC: A gene cluster containing two fungal polyketide synthases encodes the biosynthetic pathway for a polyketide, asperfuranone, in Aspergillus nidulans. J Am Chem Soc 2009, 13:2965-2970.
- [41]Bergmann S, Schümann J, Scherlach K, Lange C, Brakhage AA, Hertweck C: Genomics-driven discovery of PKS-NRPS hybrid metabolites from Aspergillus nidulans. Nat Chem Biol 2007, 3:213-217.
- [42]Gerke J, Bayram O, Feussner K, Landesfeind M, Shelest E, Feussner I, Braus GH: Breaking the silence: protein stabilization uncovers silenced biosynthetic gene clusters in the fungus Aspergillus nidulans. Appl Environ Microbiol 2012, 78:8234-8244.
- [43]Bergmann S, Funk AN, Scherlach K, Schroeckh V, Shelest E, Horn U, Hertweck C, Brakhage AA: Activation of a silent fungal polyketide biosynthesis pathway through regulatory cross talk with a cryptic nonribosomal peptide synthetase gene cluster. Appl Environ Microbiol 2010, 76:8143-8149.
- [44]Chiang YM, Szewczyk E, Davidson AD, Entwistle R, Keller NP, Wang CC, Oakley BR: Characterization of the Aspergillus nidulans monodictyphenone gene cluster. Appl Environ Microbiol 2010, 76:2067-2074.
- [45]Martin J: Clusters of genes for the biosynthesis of antibiotics: regulatory genes and overproduction of pharmaceuticals. J Ind Microbiol 1992, 9:73-90.
- [46]Brown DW, Yu JH, Kelkar HS, Fernandes M, Nesbitt TC, Keller NP, Adams TH, Leonard TJ: Twenty-five coregulated transcripts define a sterigmatocystin gene cluster in Aspergillus nidulans. Proc Natl Acad Sci USA 1996, 93:1418-1422.
- [47]Bok JW, Hoffmeister D, Maggio-Hall LA, Murillo R, Glasner JD, Keller NP: Genomic mining for Aspergillus natural products. Chem Biol 2006, 13:31-37.
- [48]Robinson SL, Panaccione DG: Chemotypic and genotypic diversity in the ergot alkaloid pathway of Aspergillus fumigatus. Mycologia 2012, 104:804-812.
- [49]Maiya S, Grundmann A, Li SM, Turner G: The fumitremorgin gene cluster of Aspergillus fumigatus: identification of a gene encoding brevianamide F synthetase. ChemBioChem 2006, 7:1062-1069.
- [50]Gardiner DM, Howlett BJ: Bioinformatic and expression analysis of the putative gliotoxin biosynthetic gene cluster of Aspergillus fumigatus. FEMS Microbiol Lett 2005, 248:241-248.
- [51]Crabtree J, Angiuoli SV, Wortman JR, White OR: Sybil: methods and software for multiple genome comparison and visualization. Meth Mol Biol 2007, 408:93-108.
- [52]Bok JW, Chiang YM, Szewczyk E, Reyes-Dominguez Y, Davidson AD, Sanchez JF, Lo HC, Watanabe K, Strauss J, Oakley BR, Wang CC, Keller NP: Chromatin-level regulation of biosynthetic gene clusters. Nat Chem Biol 2009, 5:462-464.
- [53]de Groot PW, Brandt BW, Horiuchi H, Ram AF, de Koster CG, Klis FM: Comprehensive genomic analysis of cell wall genes in Aspergillus nidulans. Fungal Genet Biol 2009, 46:S72-81.
- [54]Remm M, Storm CE, Sonnhammer EL: Automatic clustering of orthologs and in-paralogs from pairwise species comparisons. J Mol Biol 2001, 314:1041-1052.
- [55]Altenhoff AM, Dessimoz C: Phylogenetic and Functional Assessment of Orthologs Inference Projects and Methods. PLoS Comput Biol 2009, 5:e1000262.
- [56]Zdobnov EM, Apweiler R: InterProScan–an integration platform for the signature-recognition methods in InterPro. Bioinformatics 2001, 17:847-848.