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Molecular and Cellular Biology, September 1998, p. 5052-5061, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
An Exposed KID-Like Domain in Human T-Cell
Lymphotropic Virus Type 1 Tax Is Responsible for the Recruitment
of Coactivators CBP/p300
Robert
Harrod,1
Yong
Tang,1
Christophe
Nicot,1
Hsieng S.
Lu,2
Alex
Vassilev,3
Yoshihiro
Nakatani,3 and
Chou-Zen
Giam1,*
Department of Microbiology and Immunology,
Uniformed Services University of the Health Sciences, Bethesda,
Maryland 208141;
Department of Protein
Structure, Amgen Inc., Thousand Oaks, California
913202; and
Laboratory of Molecular
Growth Regulation, National Institute of Child Health and Human
Development, National Institutes of Health, Bethesda, Maryland
20892-27533
Received 7 April 1998/Returned for modification 18 May
1998/Accepted 9 June 1998
Human T-cell lymphotropic virus type 1 (HTLV-1) transcriptional
activation is mediated by the viral transactivator, Tax, and three
21-bp repeats (Tax response element [TxRE]) located in the U3 region
of the viral long terminal repeat (LTR). Each TxRE contains a core
cyclic AMP response element (CRE) flanked by 5' G-rich and 3' C-rich
sequences. The TxRE binds CREB (CRE-binding protein) and Tax to form a
ternary complex and confers Tax-dependent transactivation. Recent data
indicate that Tax functions as a specific link to connect CREB-binding
protein (CBP)/p300 in a phosphorylation-independent manner to
CREB/ATF-1 assembled on the viral 21-bp repeats. Glutathione S-transferase pull-down performed with Tax deletion mutants
and peptide competition have localized the site in Tax critical for binding CBP/p300 to a highly protease-sensitive region around amino
acid residues 81 to 95 (81QRTSKTLKVLTPPIT95)
which lies between the domains previously proposed to be important for
CREB binding and Tax subunit dimerization. Amino acid residues around the trypsin- and chymotrypsin-sensitive sites
(88KVL90) of Tax bear resemblance to those in
the kinase-inducible domain of CREB (129SRRPSYRKILNE140) surrounding Ser-133, which
undergoes signal-induced phosphorylation to recruit CBP/p300.
Site-directed mutagenesis of residues in this domain (R82A, K85A, K88A,
and V89A) resulted in proteins which failed to transactivate from the
HTLV-1 LTR in vivo. These mutants (K85A, K88A, and V89A) bind CREB with
similar affinities as wild-type Tax, yet interaction with CBP/p300 is abrogated in various biochemical assays, indicating that the
recruitment of CBP/p300 is crucial for Tax transactivation. A Tax
mutant, M47, defective in the COOH-terminal transactivation domain,
continued to interact with CBP/p300, suggesting that interactions with
additional cellular factors are required for proper Tax function.
*
Corresponding author. Mailing address: Department of
Microbiology and Immunology, Uniformed Services University of the
Health Sciences, 4301 Jones Bridge Rd., Bethesda, MD 20814. Phone:
(301) 295-9624. Fax: (301) 295-1545. E-mail:
giam{at}bob.usuf2.usuhs.mil.
Molecular and Cellular Biology, September 1998, p. 5052-5061, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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