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. Author manuscript; available in PMC: 2009 May 12.
Published in final edited form as: Cell Host Microbe. 2008 Mar 13;3(3):158–167. doi: 10.1016/j.chom.2008.02.002

Figure 4.

Figure 4

Tat binds to the acetyl lysine-binding site in SIRT1

(A) Coimmunoprecipitation assay of Tat and SIRT1 deletion mutants. Expression vectors for Myc-tagged murine Sir2α deletion mutants or full length human SIRT1 (each 1 μg) were transiently cotransfected with constructs expressing FLAG-tagged Tat (1 μg) in 293 cells. Immunoprecipitations were performed with α-FLAG agarose and western blotting with α-Myc and α-FLAG antibodies.

(B) Schematic summary of Tat binding to murine Sir2α deletion mutants. The relative localization of the human SIRT1 HDAC domain is shown at the bottom.

(C) Schematic representation of the human SIRT1 HDAC domain with point mutations.

(D) In vitro HDAC assays of immunoprecipitated human SIRT1 mutants with and without NAD+.

(E) Coimmunoprecipitation of SIRT1 mutants and Tat in 293 cells. Cell extracts were immunoprecipitated with α-T7 agarose followed by western blotting with α-FLAG and α-T7 antibodies.