FIG 1.

PELP1 interacts with the macrodomain of the histone variant macroH2A1.1 in an ADPR-independent manner. (A) Schematic diagram of the histone variant macroH2A1.1 and GST-tagged macrodomain. The histone H2A-like region (H2A), macrodomain, and the basic linker region are shown. The GST-macro1.1 fusion consists of GST fused to the nonhistone regions only. GST alone, used as a control, is shown for comparison. (B) Silver-stained image of GST-macro1.1-interacting proteins from a GST pulldown assay. HeLa nuclear extract (NE) was added as indicated. GST-macro1.1-specific protein bands were excised from the gel and identified by mass spectrometry. Macrodomain-interacting proteins, PELP1 and PARP-1, are shown. The full data set of identified proteins can be found in Fig. S3 and Table S1 in the supplemental material. (C) Western blots for PARP-1 and PELP1 from GST pulldown assay as described for panel B. ADPR or NAD⁺ at 20 μM was added as indicated.