FIG. 7.

EBNA1 domains involved in PARylation and PAR binding. (A) Purified GST-EBNA1 fusion proteins were assayed for in vitro poly(ADP-ribosyl)ation in the presence of NAD (1 mM) and then analyzed by SDS-PAGE and Western blotting with anti-PAR, anti-GST or anti-PARP1 antibody. (B) Purified GST-EBNA1 fusion proteins were assayed as above in the absence of NAD. (C) Purified GST, GST-LR1, or GST-LR2 protein was incubated with ³²P-labeled PAR and assayed by EMSA. (D) Schematic of EBNA1 protein domains. Linking regions 1 and 2 (LR1 and LR2), glycine-alanine repeats (GAr), nuclear localization sequence (NLS), and the DNA binding domain (DBD) are indicated. PAR acceptor sites are indicated. (D) Schematic summary of EBNA1 domains involved in PARylation and PAR binding as determined in panels A to C.