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Sir2p Associates with the Sum1 Complex.

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posted on 21.02.2013, 12:19 by Meleah A Hickman, Laura N Rusche

(A) Association of HA-Sir2p (pRO298) with the DTR1 promoter in wild-type (W3031-a), hst1-N291A (LRY1306), and hst1Δ (LRY198) strain backgrounds was assessed by ChIP followed by quantitative PCR. Also shown is the association of Hst1p-HA (LRY558) with the same promoters in a wild-type background. Association at the DTR1 (B primers) promoter was normalized to an internal control, the ATS1 promoter, which is not regulated by Hst1p or Sir2p. The y-axis represents the enrichment of DTR1 relative to ATS1. A value of one (dashed line) corresponds to no enrichment.

(B) The association of HA-Sir2p or Hst1p-HA with Sum1p was assessed by co-precipitation. Sum1p was immunoprecipitated from whole-cell extracts from the same strains used in (A), and the precipitated material was examined by immunoblotting with an α-mouse HA antibody to detect Hst1p-HA or HA-Sir2p.

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