1. Comparative genomics of fungal cytochrome P450s David R. Nelson University of Tennessee, Memphis Nectria haematococca Aspergillus fumigatus Aspergillus nidulans Fusarium oxysporum Aspergillus oryzae N. crassa Fusarium graminearum

2. CYP4 CYP2 CYP6 CYP9 CYP3 CYP1 Animal P450 Abundance by Family 110 Families 38% of animal P450 families have only 1 or 2 sequences. 2740 named animal P450s

3. 0 20 40 60 80 100 120 470 Number of P450 genes and pseudogenes Plant P450 Abundance by Family 95 Families CYP71 CYP81 CYP76 CYP72 CYP73 CYP74 Only 28% of Plant P450 Families have 1 or 2 members 2867 named plant P450s

4. 60% of fungal P450 families have only 1 or 2 sequences. CYP51 CYP52 CYP61 CYP65 1384 named fungal CYPs 329 families

5. Prokaryote P450 Abundance by Family 61% of prokaryote P450 families have only 1 or 2 sequences. CYP107 CYP105 CYP153 CYP110 CYP102 205 families 813 named bacterial CYPs

6. Fungal P450 clans defined as the deepest branches on phylogenetic trees.

7. Deng J., Carbone I. and Dean R.A. The evolutionary history of cytochrome P450 genes in four filamentous ascomycetes. BMC Evolutionary Biology 2007, 7:30 Doi:10.1186/1471-2148-7-30 Fusarium graminearum Neurospora crassa Magnaporthe grisea Aspergillus nidulans

8. Asp. ochraceous Fus. sporotrichiodes Fus. verticilloides Fus. oxysporum Asp. parasiticus Asp. terreus Asp. clavatus Asp. flavus Neo. fischeri Asp. fumigatus Mag. grisea Neurospora crassa Asp. oryzae Asp. niger Asp. nidulans Fus. graminearum Nectria haematococca Asp. ochraceous Fus. sporotrichiodes Fus. verticilloides Fus. oxysporum Asp. parasiticus Asp. terreus Asp. clavatus Asp. flavus Neo. fischeri Asp. fumigatus Mag. grisea Neurospora crassa Asp. oryzae Asp. niger Asp. nidulans Fus. graminearum Nectria haematococca

9. Asp. ochraceous Fus. sporotrichiodes Fus. verticilloides Fus. oxysporum Asp. parasiticus Asp. terreus Asp. clavatus Asp. flavus Neo. fischeri Asp. fumigatus Mag. grisea Neurospora crassa Asp. oryzae Asp. niger Asp. nidulans Fus. graminearum Nectria haematococca Asp. ochraceous Fus. sporotrichiodes Fus. verticilloides Fus. oxysporum Asp. parasiticus Asp. terreus Asp. clavatus Asp. flavus Neo. fischeri Asp. fumigatus Mag. grisea Neurospora crassa Asp. oryzae Asp. niger Asp. nidulans Fus. graminearum Nectria haematococca

10. Old familiesNew families

11. The 52 most widely distributed P450 families among 8 completed filamentous fungal genomes benzoate-para-hydroxylase ergosterol biosynthesis fatty acid hydroxylase multiple toxins alkane/ fatty acid hydroxylase phenylacetate 2-hydroxylase aflatoxin/trichothecene nitric oxide reductase sterigmatocystin synthesis gliotoxin biosynthesis pigment biosynthesis

12. The distribution of 252 P450 families across 8 completely sequenced filamentous fungal genomes

13. Vertebrate cars: all families standard, some subfamilies optional

14. Fungal cars: CYP51, CYP61 standard, 250 other families optional

15. R. Perrin, N. Federova, J.-W. Bok, R.A. Cramer Jr., J. Wortman, H. Kim, W. Nierman and N. Keller, Transcriptional regulation of chemical diversity in Aspergillus fumigatus by LaeA. PLoS Pathogen 3 (2007), p. e50. 22 gene clusters were identified. 9 of these clusters contained 19 P450 genes. 38% of the P450s in Aspergillus fumigatus showed differential expression in a delta LaeA strain.

16. The epipolythiodioxopiperazine (ETP) class of fungal toxins: distribution, mode of action, functions and biosynthesis Donald M. Gardiner, Paul Waring and Barbara J. Howlett Microbiology 151 (2005), 1021-1032 CYP5082B1CYP5093A1 CYP613B1 CYP5085A1 CYP566C2 geranylgeranylDP synthase (Also in Neosartorya fischeri)

17. Aspergillus nidulans P450s in chromosome order

18. Chromosome 12 P450 gene cluster b (249111-271400) 578A2 5040A1 532J1 504B4 Homogentisate 1,2 diooxygenase Fumarylacetoacetate hydrolase 3,4-dihydroxyphenylacetate 6-hydroxylase This is part of the phenylacetate catabolic pathway that includes 504B4, 504A6, Homogentisate 1,2 diooxygenase and Fumarylacetoacetate hydrolase. The 504A6 gene is 60kb downstream of 504B4

19. Aspergillus nidulans P450s in chromosome order

20. The epipolythiodioxopiperazine (ETP) class of fungal toxins: distribution, mode of action, functions and biosynthesis Donald M. Gardiner, Paul Waring and Barbara J. Howlett Microbiology 151 (2005), 1021-1032 CYP5082B1CYP5093A1 CYP613B1 CYP5085A1 CYP566C2 geranylgeranylDP synthase (Also in Neosartorya fischeri)

21. Aspergillus nidulans P450s in chromosome order

22. A. nidulans A. clavatus A. oryzae A. terreus 3269 3270 3271 3272 3273 3274 3275 3276 3277 3278 3279 3280 3281 ZnCysMFS667A1PKS654A1567E1 Terpene cylase 567D1 3839 3840 3841 3842 3843 3844 3845 3846 3847 3848 3849 3850 567D3ZnCys Terpene cylase 567E654APKS667A2 ZnCys 0097 0098 0099 0100 0101 0102 0103 Terpene cylase 654B1567D2 MFS 682B2 Terpene cylase

23. Aspergillus nidulans P450s in chromosome order