1. Inhibition of Retrovirus Replication by Human Schlafen 11

2. Murine Schlafen (Slfn) Family Slfn5 Slfn8 Slfn9 Putative AAA domain Slfn14 Slfn3 Slfn4 Slfn10 Slfn1 Slfn2 In search for IRF3-regulated genes, we found multiple members of the Schlafen (Slfn) gene family… Putative RNA helicase domain

3. Human Slfn Family Slfn5 Slfn11 Slfn13 Slfn12 Slfn12L Putative AAA domain Slfn14 Putative RNA helicase domain

4. Discovery of Slfns The Schlafen (Slfn) family of genes are differentially regulated during thymocyte maturation and are preferentially expressed in the lymphoid tissues. Ectopic expression of the prototype member Slfn1 early in the T lineage alters cell growth and development. Expression of Slfn1 in fibroblasts and thymoma cells either retards or ablates cell growth. Slfn1 knockout mice had no significant phenotype

5. Transgenic T-cell specific expression of muSlfn8 in mice resulted in impaired T cell development (Geserick et al., Int Immunol. 2004) Mutation of muSlfn2 causes loss of immune quiescence in T cells and monocytes (Berger et al., Nature Immun. 2010) muSlfn1 causes a cell cycle arrest by inhibiting induction of cyclin D1 (Brady et al., JBC 2005) Role of Schlafen 2 (SLFN2) in the generation of interferon alpha-induced growth inhibitory responses (Katsoulidis et al., JBC 2009) Lack of reproducible growth inhibition by muSlfn1 and muSlfn2 in vitro (Zhao et al., Blood Cells Mol Dis. 2008) In the next 12 years…

6. AAA-domain Snider et al. Genome Biology 2008 9:216 membrane fusion protein translocation ER-associated degradation proteolysis peroxisome biogenesis mitosis, meiosis apoptosis microtubule dynamics motors DNA replication DNA recombination and repair transcriptional regulation homo/hetero-multimer formation “ATPases Associated with various cellular Activities”

7. mRNA Levels of Slfns in Human Fibroblasts (HFF) upon Stimulation: Slfn expression profiles mRNA Levels of Slfns in Cell lines:

8. huSlfn11 knock-down HEK293 cell lines Slfns are induced by Interferons and TLR ligands Putative helicase domain (similarity to RIG-I ??) => Do they function as pathogen sensors or activators of innate immune responses?

9. Retroviral Replication Cycle

10. huSlfn11 does not alter Infection by MSCV or HIV MSCV HIV

11. HIV Infection and Replication Cycle Do Slfns act as effectors in the innate immune response?

12. huSlfn11 restricts production of MSCV or HIV MSCV HIV

13. Slfn5 (891 a.a.) RNA helicase Divergent AAA domain Slfn11 (901 a.a.) Slfn11N (1-579 a.a.) Slfn11C (533-901 a.a.) Truncation mutants of huSlfn11

14. huSlfn11N restricts production of MSCV or HIV MSCV HIV

15. huSlfn11 does not restrict production of AAV or HSV

16. Mechanism? Extracellular p24 and vRNA levels correlate with viral titers by infection assay => Fewer viral particles are produced

17. Virus budding & release are not affected by huSlfn11

18. Intracellular vRNA levels are not affected by huSlfn11

19. huSlfn11 specifically inhibits viral protein synthesis

20. huSlfn11 appears to inhibit viral protein translation by interference with -1 frameshifting

21. CEM cells and HIV LAI

22. David Lab Manqing Li Hilary Sandig Elaine Kao Kirsten Vroom Irene Pedersen Dennis Otero Hiroshi Moro Yoshinari Tanabe Takeaki Nishibori Salk Institute Matt Weitzmann Sebastian Landry John Young THANKS... UCSD-CFAR John Guatelli Davey Smith