Transcription factor binding sites in the pol gene intragenic regulatory region of HIV-1 are important for virus infectivity

被引:24
作者
Goffin, W
Demonté, D
Vanhulle, C
de Walque, S
de Launoit, Y
Burny, A
Collette, Y
Van Lint, C
机构
[1] Univ Libre Bruxelles, Serv Chim Biol, Inst Biol & Med Mol, Mol Virol Lab, B-6041 Gosselies, Belgium
[2] Free Univ Brussels, Mol Virol Lab, Fac Med, B-1070 Brussels, Belgium
[3] Univ Lille 1, CNRS, Inst Pasteur Lille, Inst Biol Lille,UMR 8117, F-59021 Lille, France
[4] INSERM, U119, F-13009 Marseille, France
关键词
D O I
10.1093/nar/gki720
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have previously identified in the pol gene of human immunodeficiency virus type 1 (HIV-1) a new positive transcriptional regulatory element (nt 4481-4982) containing recognition sites for nuclear proteins (sites B, C, D and a GC-box) [C. Van Lint, J. Ghysdael, P. Paras, Jr, A. Burny and E. Verdin (1994) J. Virol 68, 2632-2648]. In this study, we have further physically characterized each binding site and have shown that the transcription factors Oct-1, Oct-2, PU.1, Sp1 and Sp3 interact in vitro with the pol region. Chromatin immunoprecipitation assays using HIV-infected cell lines demonstrated in the context of chromatin that Sp1, Sp3, Oct-1 and PU.1 are recruited to the HS7 region in vivo. For each site, we have identified mutations abolishing factor binding to their cognate DNA sequences without altering the underlying amino acid sequence of the integrase. By transient transfection assays, we have demonstrated the involvement of the pol binding sites in the transcriptional enhancing activity of the intragenic region. Our functional results with multimerized wild-type and mutated pol binding sites separately (i.e. in the absence of the other sites) have demonstrated that the PU.1, Sp1, Sp3 and Oct-1 transcription factors regulate the transcriptional activity of a heterologous promoter through their respective HS7 binding sites. Finally, we have investigated the physiological role of the HS7 binding sites in HIV-1 replication and have shown that these sites are important for viral infectivity.
引用
收藏
页码:4285 / 4310
页数:26
相关论文
共 74 条
[1]   Selective regulation of human immunodeficiency virus-infected CD4+ lymphocytes by a synthetic immunomodulator leads to potent virus suppression in vitro and in hu-PBL-SCID mice [J].
Bahr, GM ;
Darcissac, ECA ;
Castéran, N ;
Amiel, C ;
Cocude, C ;
Truong, MJ ;
Dewulf, J ;
Capron, A ;
Mouton, Y .
JOURNAL OF VIROLOGY, 2001, 75 (15) :6941-6952
[2]   ISOLATION OF A T-LYMPHOTROPIC RETROVIRUS FROM A PATIENT AT RISK FOR ACQUIRED IMMUNE-DEFICIENCY SYNDROME (AIDS) [J].
BARRESINOUSSI, F ;
CHERMANN, JC ;
REY, F ;
NUGEYRE, MT ;
CHAMARET, S ;
GRUEST, J ;
DAUGUET, C ;
AXLERBLIN, C ;
VEZINETBRUN, F ;
ROUZIOUX, C ;
ROZENBAUM, W ;
MONTAGNIER, L .
SCIENCE, 1983, 220 (4599) :868-871
[3]   THE PRODUCT OF THE C-ETS-1 PROTOONCOGENE AND THE RELATED ETS2 PROTEIN ACT AS TRANSCRIPTIONAL ACTIVATORS OF THE LONG TERMINAL REPEAT OF HUMAN T-CELL LEUKEMIA-VIRUS HTLV-1 [J].
BOSSELUT, R ;
DUVALL, JF ;
GEGONNE, A ;
BAILLY, M ;
HEMAR, A ;
BRADY, J ;
GHYSDAEL, J .
EMBO JOURNAL, 1990, 9 (10) :3137-3144
[4]   Regulation of the activity of Sp1-related transcription factors [J].
Bouwman, P ;
Philipsen, S .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 2002, 195 (1-2) :27-38
[5]   Transcription factor Sp3 is essential for post-natal survival and late tooth development [J].
Bouwman, P ;
Göllner, H ;
Elsässer, HP ;
Eckhoff, G ;
Karis, A ;
Grosveld, F ;
Philipsen, S ;
Suske, G .
EMBO JOURNAL, 2000, 19 (04) :655-661
[6]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[7]   A prominent role for Sp1 during lipopolysaccharide-mediated induction of the IL-10 promoter in macrophages [J].
Brightbill, HD ;
Plevy, SE ;
Modlin, RL ;
Smale, ST .
JOURNAL OF IMMUNOLOGY, 2000, 164 (04) :1940-1951
[8]   Upstream stimulatory factors binding to an E box motif in the R region of the bovine leukemia virus long terminal repeat stimulates viral gene expression [J].
Calomme, C ;
Nguyên, TLA ;
de Launoit, Y ;
Kiermer, V ;
Droogmans, L ;
Burny, A ;
Van Lint, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (11) :8775-8789
[9]   TISSUE-SPECIFIC TRANSCRIPTION PREFERENCE AS A DETERMINANT OF CELL TROPISM AND LEUKEMOGENIC POTENTIAL OF MURINE RETROVIRUSES [J].
CELANDER, D ;
HASELTINE, WA .
NATURE, 1984, 312 (5990) :159-162
[10]   OCTAMER-BINDING PROTEINS IN DIVERSE HEMATOPOIETIC-CELLS [J].
COCKERILL, PN ;
KLINKEN, SP .
MOLECULAR AND CELLULAR BIOLOGY, 1990, 10 (03) :1293-1296