Role of SUMO-1 and SUMO interacting motifs in rhesus TRIM5α-mediated restriction
- Equal contributors
1 Department of Microbiology and Immunology Stritch School of Medicine, Loyola University Chicago, 2160 S. 1st Ave. 508, Building 102, Room 5651, Maywood, IL, 60153, USA
2 Department of Biochemistry and Molecular Biophysics, Columbia University, 701W168th street, HHSC1310, New York, NY 10032USA
3 Howard Hughes Medical institute
4 Departamento de Ciencias Biologicas, Facultad de Ciencias Biologicas, Universidad Andres Bello, Los Fresnos 52, Viña del Mar, Chile
Retrovirology 2013, 10:10 doi:10.1186/1742-4690-10-10Published: 1 February 2013
TRIM5α is a member of the tripartite motif family of proteins that restricts retroviral infection in a species-specific manner. The restriction requires an interaction between the viral capsid lattice and the B30.2/SPRY domain of TRIM5α. Previously, we determined that two SUMO interacting motifs (SIMs) present in the B30.2/SPRY domain of human TRIM5α (huTRIM5α) were important for the restriction of N-tropic Murine Leukemia Virus. Here, we examined whether SUMO expression and the SIM1 and SIM2 motifs in rhesus monkey TRIM5α (rhTRIM5α) are similarly important for Human Immunodeficiency Type 1 (HIV-) restriction.
We found that mutation of SIM1 and SIM2 of rhTRIM5α abolished the restriction of HIV-1 virus. Further, knockdown of SUMO-1 in rhTRIM5α expressing cells abolished restriction of HIV-1. These results may be due, in part, to the ability of SUMO-1 to stabilize rhTRIM5α protein expression, as SUMO-1 knockdown increased rhTRIM5α turnover and the mutations in SIM1 and SIM2 led to more rapid degradation than the wild type protein. The NF-κB signaling ability of rhTRIM5α was also attenuated by SUMO-1 knockdown. Finally, upon inhibition of CRM1-dependent nuclear export with Leptomycin B (LMB), wild type rhTRIM5α localized to SUMO-1 bodies in the nucleus, while the SIM1 and SIM2 mutants did not localize to SUMO-1.
Our results suggest that the rhTRIM5α B30.2/SPRY domain is not only important for the recognition of the HIV-1 CA, but it is also important for its association with SUMO-1 or SUMO-1 modified proteins. These interactions help to maintain TRIM5α protein levels and its nuclear localization into specific nuclear bodies.