STRUCTURAL AND FUNCTIONAL ANALYSES OF THE PROMOTER OF THE MURINE MULTIDRUG RESISTANCE GENE MDR3/MDRLA REVEAL A NEGATIVE ELEMENT CONTAINING THE AP-1 BINDING-SITE

被引:46
作者
IKEGUCHI, M [1 ]
TEETER, LD [1 ]
ECKERSBERG, T [1 ]
GANAPATHI, R [1 ]
KUO, MT [1 ]
机构
[1] CLEVELAND CLIN FDN, RES INST, CLEVELAND, OH 44195 USA
关键词
D O I
10.1089/dna.1991.10.639
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 shown that the multidrug-resistance/P-glycoprotein gene, mdr3/mdr1a, is activated in mouse hepatocellular carcinomas (HCC). In this study, we show that in a number of HCC-derived cell lines (Hepa1c1c, Hepa1c1c-BprC1, and Hepa1-6) mdr3 is expressed at high levels. To investigate transcriptional regulation of mdr3 in these cells, we have isolated a DNA fragment containing the 5' portion of the mouse mdr3 gene and performed a functional analysis of its promoter. Transient transfection assays using various lengths of the promoter sequence to direct expression of the chloramphenicol acetyltransferase (CAT) reporter gene revealed that the sequence located -94 nucleotides upstream from mouse mdr3 transcription start site functions as a negative element in mouse hepatoma cells. A canonical AP-1 binding sequence TGA-GTCA located at -117 is at least in part responsible for the negative effect from the following observations: (i) Alteration of this AP-1 sequence by site-directed mutagenesis enhanced CAT expression. (ii) Expression of CAT reporter gene was elevated when double-stranded DNA containing the AP-1 sequence, but not mutated sequences, was used as a competitor in cotransfection experiment. (iii) Enhancement of the CAT expression was also seen in cotransfection experiments using recombinant plasmid DNA expressing the c-jun/c-fos proteins, which interact with AP-1 sequences. Interestingly, the proximal region of the hamster pgp1 promoter shares striking sequence similarity with that of the mouse mdr3 gene, including the AP-1 site, but the AP-1 site in the hamster promoter serves as a positive regulator. Although previous studies have demonstrated that positive and negative transcription factors can modulate gene expression through interactions with c-jun/c-fos, this is the first study to show that an AP-1 site functions as a negative cis-element in the regulation of gene expression.
引用
收藏
页码:639 / 649
页数:11
相关论文
共 41 条
  • [1] THE JUN PROTO-ONCOGENE IS POSITIVELY AUTOREGULATED BY ITS PRODUCT, JUN/AP-1
    ANGEL, P
    HATTORI, K
    SMEAL, T
    KARIN, M
    [J]. CELL, 1988, 55 (05) : 875 - 885
  • [2] PHORBOL ESTER INDUCIBLE GENES CONTAIN A COMMON CIS ELEMENT RECOGNIZED BY A TPA-MODULATED TRANS-ACTING FACTOR
    ANGEL, P
    IMAGAWA, M
    CHIU, R
    STEIN, B
    IMBRA, RJ
    RAHMSDORF, HJ
    JONAT, C
    HERRLICH, P
    KARIN, M
    [J]. CELL, 1987, 49 (06) : 729 - 739
  • [3] IP-1 - A DOMINANT INHIBITOR OF FOS/JUN WHOSE ACTIVITY IS MODULATED BY PHOSPHORYLATION
    AUWERX, J
    SASSONECORSI, P
    [J]. CELL, 1991, 64 (05) : 983 - 993
  • [4] V-SRC AND EJ RAS ALLEVIATE REPRESSION OF C-JUN BY A CELL-SPECIFIC INHIBITOR
    BAICHWAL, VR
    PARK, A
    TJIAN, R
    [J]. NATURE, 1991, 352 (6331) : 165 - 168
  • [5] CONTROL OF C-JUN ACTIVITY BY INTERACTION OF A CELL-SPECIFIC INHIBITOR WITH REGULATORY DOMAIN-DELTA - DIFFERENCES BETWEEN V-JUN AND C-JUN
    BAICHWAL, VR
    TJIAN, R
    [J]. CELL, 1990, 63 (04) : 815 - 825
  • [6] BERNHARD HP, 1973, DEV BIOL, V35, P83, DOI 10.1016/0012-1606(73)90008-0
  • [7] BRADLEY G, 1989, CANCER RES, V49, P2790
  • [8] IMMUNOHISTOCHEMICAL DETECTION OF P-GLYCOPROTEIN - PROGNOSTIC CORRELATION IN SOFT-TISSUE SARCOMA OF CHILDHOOD
    CHAN, HSL
    THORNER, PS
    HADDAD, G
    LING, V
    [J]. JOURNAL OF CLINICAL ONCOLOGY, 1990, 8 (04) : 689 - 704
  • [9] JUN-B DIFFERS IN ITS BIOLOGICAL PROPERTIES FROM, AND IS A NEGATIVE REGULATOR OF, C-JUN
    CHIU, R
    ANGEL, P
    KARIN, M
    [J]. CELL, 1989, 59 (06) : 979 - 986
  • [10] AN ALTERED PATTERN OF CROSS-RESISTANCE IN MULTIDRUG-RESISTANT HUMAN-CELLS RESULTS FROM SPONTANEOUS MUTATIONS IN THE MDR1 (P-GLYCOPROTEIN) GENE
    CHOI, K
    CHEN, C
    KRIEGLER, M
    RONINSON, IB
    [J]. CELL, 1988, 53 (04) : 519 - 529