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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
-1
subunit,
-tubulin, and
-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,
-tubulin, and
-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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