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Journal of Virology, March 2001, p. 2475-2481, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2475-2481.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

EBNA-LP Associates with Cellular Proteins Including DNA-PK and HA95

Innoc Han,1 Shizuko Harada,1 David Weaver,2 Yong Xue,1 William Lane,3 Sigurd Orstavik,4 Bjorn Skalhegg,4 and Elliott Kieff1,*

Channing Laboratory, Harvard Medical School, Boston,1 and Center for Blood Research, Harvard University,2 and Harvard Microchemistry Facility,3 Cambridge, Massachusetts, and Institute of Medical Biochemistry, University of Oslo, Blindern, N-0317 Oslo, Norway4

Received 12 September 2000/Accepted 13 November 2000

EBNA-LP-associated proteins were identified by sequencing proteins that immunoprecipitated 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), HA95, Hsp27, prolyl 4-hydroxylase alpha -1 subunit, alpha -tubulin, and beta -tubulin were identified. The fraction of total cellular HA95 that associated with FLP was very high, while progressively lower fractions of the total DNA-PKcs, Hsp70, Hsp 27, alpha -tubulin, and beta -tubulin specifically associated with EBNA-LP as determined by immunoblotting with antibodies to these proteins. EBNA-LP bound to two domains in the DNA-PKcs C terminus and DNA-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 DNA-PKcs.


* Corresponding author. Mailing address: Channing Laboratory, Harvard Medical School, 181 Longwood Ave., Boston, MA 02445. Phone: (617) 525-4252. Fax: (617) 525-4257. E-mail: ekieff{at}rics.bwh.harvard.edu.


Journal of Virology, March 2001, p. 2475-2481, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2475-2481.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.



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