EBNA-LP associates with cellular proteins including DNA-PK and HA95

EBNA-LP-associated proteins were identified by sequencing proteins that imm unoprecipitated with Flag epitope-tagged EBNA-LP (FLP) from lymphoblasts in which FLP was stably expressed. The association of EBNA-LP with Hsp70 (72/ 73) was confirmed, and sequences of DNA-PK catalytic subunit (DNA-PKcs), HA 95, Hsp27, prolyl 4-hydroxylase alpha -1 subunit, alpha -tubulin, and beta -tubulin were identified. The fraction of total cellular HA95 that associat ed with FLP was very high, while progressively lower fractions of the total DNA-PKcs, Hsp70, Hsp 27, alpha -tubulin, and beta -tubulin specifically as sociated with EBNA-LP as determined by immunoblotting with antibodies to th ese proteins. EBNA-LP bound to two domains in the DNA-PKcs C terminus and D NA-PKcs associated with the EBNA-LP repeat domain. DNA-PKcs that was bound to EBNA-LP phosphorylated p53 or EBNA-LP in vitro, and the phosphorylation of EBNA-LP was inhibited by Wortmannin, a specific in vitro inhibitor of DN A-PKcs.