A retrovirus restriction factor TRIM5alpha is transcriptionally regulated by interferons

Abstract

TRIM5alpha is a member of tripartite motif protein family and recently identified as a restriction factor for retroviral infection in a species-specific manner. Human TRIM5alpha gene is located on chromosomal position 11p15 in a cluster with other TRIM genes including TRIM6, 21, 22, and 34. We show here that interferon (IFN) upregulates TRIM5alpha mRNA expression in HeLa and HepG2 cells by performing Northern blot analysis and quantitative real-time PCR. TRIM5alpha promoter activity was IFN inducible as confirmed by luciferase assay using a reporter plasmid that contained the 5'-flanking region of TRIM5alpha. Mutational analysis has revealed that IFNs activate TRIM5alpha promoter activity through a putative interferon-stimulated response element (ISRE). Intriguingly, another IFN-responsive protein signal transducer and activator of transcription factor 1 (STAT1) binds to the ISRE sequence as shown by electrophoretic mobility shift assay using HeLa cell extracts. We have raised a specific polyclonal antibody against TRIM5alpha and confirmed that TRIM5alpha protein expression is inducible by IFN-beta in HeLa cells. These results lead us to define that the transcription and protein synthesis of TRIM5alpha could be modulated by IFN, suggesting that TRIM5alpha may play a role in an IFN-induced antiviral state against retrovirus infection.