The HIV-1 transactivator protein Tat is a critical regulator of HIV transcription primarily enabling efficient elongation of viral transcripts. Its interactions with RNA and various host factors are regulated by ordered, transient post-translational modifications. Here, we report a novel Tat modification, monomethylation at lysine 71 (K71). We found that Lys-71 monomethylation (K71me) is catalyzed by KMT7, a methyltransferase that also targets lysine 51 (K51) in Tat. Using mass spectrometry, in vitro enzymology, and modification-specific antibodies, we found that KMT7 monomethylates both Lys-71 and Lys-51 in Tat. K71me is important for full Tat transactivation, as KMT7 knockdown impaired the transcriptional activity of wild type (WT) Tat but not a Tat K71R mutant. These findings underscore the role of KMT7 as an important monomethyltransferase regulating HIV transcription through Tat.
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This work was supported by the University of California San Francisco, Gladstone Institute of Virology and Immunology Center for AIDS Research, a collaboration with JT Pharma, Gladstone Institutes, CA AIDS Research Program, and Grants R01AI083139, U19AI096113, T32IA7334-26, and P30AI027763 from the National Institutes of Health, and Grant ID F13-GI-316 from the California HIV/AIDS Research Program (CHRP).
This article contains supplemental Tables S-1 and S-2.
Ali, Ibraheem; Ramage, Holly; Boehm, Daniela; Dirk, Lynnette M. A.; Sakane, Naoki; Hanada, Kazuki; Pagans, Sara; Kaehlcke, Katrin; Aull, Katherine; Weinberger, Leor; Trievel, Raymond; Schnoelzer, Martina; Kamada, Masafumi; Houtz, Robert L.; and Ott, Melanie, "The HIV-1 Tat Protein is Monomethylated at Lysine 71 by the Lysine Methyltransferase KMT7" (2016). Horticulture Faculty Publications. 30.