1. Global Control of Aspergillus fumigatus Toxins Nancy Keller University of Wisconsin

2. Aspergillus secondary metabolites

3. Aspergillus laeA: transcriptional regulator of secondary metabolite genes Bok and Keller, Euk Cell, 3: 527-535 ST WT laeA A. nidulans GT WT laeA comp A. fumigatus

4. Time (Days) 02468 0 20 40 60 80 100 laeA Wild type 02468 20 40 60 80 100 02468 20 40 60 80 100 Survival % 02468 20 40 60 80 10 0 Complemented Mouse survival LaeA is a virulence factor in Aspergillus fumigatus Bok et al, Euk Cell, 4:1574-1582 51560 0 10 20 30 40 50 60 WT laeA Incubation (minutes) PMN death (%) Macrophage phagocytosis WT laeA Neutrophil death

6. Gardiner and Howlett 2005, Microbiology

7. Number of conidia (10 6 /ml/cm 2 ) 0 10 20 30 40 50 60 WT gliZ gliZ + gliZ A. fumigatus gliZ grows at same rate, produces same number of spores

8. Gt WT gliZ + gliZ gliZ + gliZ laeA gliZ does not produce gliotoxin

9. gliZ regulates other genes in the gliotoxin gene cluster gliZ rRNA 36hr 72hr WT laeA gliZ gliZcom gliI A B 0.5 kb 1.0 kb 1 2 3 4

10. Murine pulmonary model

11. Gliotoxin standard Non-infected lung gliZ laeA WT 1000 5.005.205.405.605.806.006.206.406.606.807.007.20 Time (min) 0 0 17 0 520 0 19 0 312 0 92 5.87 5.80 5.87 Gliotoxin chromatograms lung tissue gliZ + gliZ

12. A B p=0.04 0 5 10 15 20 25 30 35 40 45 WT laeA gliZ gliZcom8.3 PI + (% PMNs) 0 10 20 30 40 50 60 Annexin + (% PMNs) WT laeA gliZ gliZcom8.3

13. Acknowledgements Keller Lab –Dr. JinWoo Bok –Elyse Bolterstein –Dr. Marion Brodhagen –DaWoon Chung –Tom Hammond –Lane Milde –Dr. Robyn Perrin –James Scott –E. Keats Shwab Collaborators –Drs. Bill Nierman, Natalie Fedorova The Institute for Genomic Research (microarrays) –Dr. David Andes Department of Medicine, Section of Infectious Disease, UW Madison –Drs. Kieren Marr and Arunmozhi Balajee Fred Hutchinson Cancer Research Center –Drs. Jens Frisvad & Kristian Fog Nielsen Department of Biotechnology, Technical University of Denmark