Exon 4-encoded acidic domain in the epithelium-restricted Ets factor, ESX, confers potent transactivating capacity and binds to TATA-binding protein (TBP)

被引:25
作者
Chang, CH
Scott, GK
Baldwin, MA
Benz, CC
机构
[1] Univ Calif San Francisco, Dept Med, Div Hematol Oncol, San Francisco, CA 94143 USA
[2] Univ Calif San Francisco, Dept Neurol, San Francisco, CA 94143 USA
[3] Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA
[4] Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA
关键词
ESX; human Ets factor; genomic organization; transactivation domain;
D O I
10.1038/sj.onc.1202674
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
The Ets gene family has a complex evolutionary history with many family members known to regulate genetic programs essential for differentiation and development, and some known for their involvement in human tumorigenesis. To understand the biological properties associated with a recently described epithelium-restricted Ets factor ESX, an 11 kb fragment from the 1q32.2 genomically localized human gene was cloned and analysed. Upstream of the ESX promoter region in this genomic fragment lies the terminal exon of a newly identified gene that encodes a ubiquitin-conjugating enzyme variant, UEV-1. Tissues expressing ESX produce a primary 2.2 kb transcript along with a 4.1 kb secondary transcript arising by alternate poly(A) site selection and uniquely recognized by a genomic probe from the 3' terminal region of the 11 kb clone. Endogenous expression of ESX results in a 42 kDa nuclear protein having fivefold greater affinity for the chromatin-nuclear matrix compartment as compared to other endogenous transcription factors like AP-2 and the homologous Ets factor, ELF-1. Exon mapping of the modular structure inferred from ESX cDNA and construction of GAL4(DBD)-ESX expression constructs were used to identify a transactivating domain encoded by exon 4 having comparable potency to the acidic transactivation domain of the viral transcription factor, VP16. This exon 4-encoded 31 amino acid domain in ESX was shown by mutation and deletion analysis to possess a 13 residue acidic transactivation core which, based on modeling and circular dichroism analysis, is predicted to form an amphipathic alpha-helical secondary structure. Using recombinant GST-ESX (exon 4) fusion proteins in an in vitro pull-down assay, this ESX transactivation domain was shown to bind specifically to one component of the general transcription machinery, TATA-binding protein (TBP). Transient transfection experiments confirmed the ability of this TBP-binding transactivation domain in ESX to squelch heterologous promoters independent of any promoter binding as efficiently as the transactivation domain from VP16.
引用
收藏
页码:3682 / 3695
页数:14
相关论文
共 46 条
  • [1] The expression of a novel, epithelium-specific ets transcription factor is restricted to the most differentiated layers in the epidermis
    Andreoli, JM
    Jang, SI
    Chung, E
    Coticchia, CM
    Steinert, PM
    Markova, NG
    [J]. NUCLEIC ACIDS RESEARCH, 1997, 25 (21) : 4287 - 4295
  • [2] [Anonymous], 1989, SYNTHETIC OLIGONUCLE
  • [3] ASHAR HR, 1995, CELL, V82, P57
  • [4] Ausubel FA, 1995, CURRENT PROTOCOLS MO
  • [5] Genomic organization of TEL: The human ETS-variant gene 6
    Baens, M
    Peeters, P
    Guo, CY
    Aerssens, J
    Marynen, P
    [J]. GENOME RESEARCH, 1996, 6 (05): : 404 - 413
  • [6] EFFECTS OF PROVIRUS INTEGRATION IN THE TPL-1/ETS-1 LOCUS IN MOLONEY MURINE LEUKEMIA VIRUS-INDUCED RAT T-CELL LYMPHOMAS - LEVELS OF EXPRESSION, POLYADENYLATION, TRANSCRIPTIONAL INITIATION, AND DIFFERENTIAL SPLICING OF THE ETS-1 MESSENGER-RNA
    BELLACOSA, A
    DATTA, K
    BEAR, SE
    PATRIOTIS, C
    LAZO, PA
    COPELAND, NG
    JENKINS, NA
    TSICHLIS, PN
    [J]. JOURNAL OF VIROLOGY, 1994, 68 (04) : 2320 - 2330
  • [7] Molecular cloning and expression of Ehf, a new member of the ets transcription factor oncoprotein gene family
    Bochert, MA
    Kleinbaum, LA
    Sun, LY
    Burton, FH
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 246 (01) : 176 - 181
  • [8] PROPERTIES OF INITIATOR-ASSOCIATED TRANSCRIPTION MEDIATED BY GAL4-VP16
    CHANG, CB
    GRALLA, JD
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1993, 13 (12) : 7469 - 7475
  • [9] ESX: A structurally unique Ets overexpressed early during human breast tumorigenesis
    Chang, CH
    Scott, GK
    Kuo, WL
    Xiong, XH
    Suzdaltseva, Y
    Park, JW
    Sayre, P
    Erny, K
    Collins, C
    Gray, JW
    Benz, CC
    [J]. ONCOGENE, 1997, 14 (13) : 1617 - 1622
  • [10] Two promoters direct expression of the murine Spi-B gene, an Ets family transcription factor
    Chen, HM
    Gonzalez, DA
    Radomska, HS
    Voso, MT
    Sun, Z
    Zhang, P
    Zhang, DE
    Tenen, DG
    [J]. GENE, 1998, 207 (02) : 209 - 218