1. Lab of Biochemistry Division of Influenza viruses, Korea Center for Disease Control and Prevention Mi-Seon Kim

2. Introduction Identification method In vitro experiments In vivo experiments Conclusion Contents

3. Introduction What’s Candida albicans? – Opportunistic fungal pathogen of humans – Invasion to internal organs through the bloodstream  systemic infections – C. albicans & Mortality rate

4. Introduction Previous work – Target : Cell wall protein Why is the Cell-surface protein? – More accessible to drugs than intercellular targets Cell surface protein –ALS1 –HWP1

5. Domain organization of a typical fungal cell-wall protein Glycosylation

6. CSF4 Procedure of Identification of cell-surface proteins in C. albicans 6 CSF 1~6 RT-PCR using primers internal to the ORF 180 candidates

7. Primary sequence analysis of C.albicans Csf4 protein

8. s First allele replacement Second allele replacement First deletion cassette generated by 3-way PCR Second deletion cassette Target sequence Target sequence

9. Filamentation assay Clinical isolationParental strainMutant Invasion upon Agar Filamentation(Hyphae formation) Growth on the Agar surface Severe defect of Filamentation No filaments around colony Targeted gene = No growth factor Viable csf4 null mutant Similar growth rate (csf4 null mutant & wild type)

10. Support a role for Csf4 in the adhesion of C.albicans to mammalian cell Adherence assay Wild- type

11. Virulence Assay in vivo Csf4 null_HC SC5314 Uninfected BWP17_ARG/HC SC5314 Csf4 null_HC als1 null_HC

12. CSF4 – Factor of Rapid invasion – Destrction of key organ Virulence Assay in vivo

13. Conclusion Cell-surface proteins represent very attractive targets Csf4 belongs to the familly of yeast glycosidases Csf4 is important for adhesion of C.albicans to mammalian cells in vitro Csf4 is good candidates for new virulence factor Csf4 is represents a very promising target for the development of antifungal drugs

14. ★ Thank you