Cell
Volume 52, Issue 5, 11 March 1988, Pages 723-729
Journal home page for Cell

Definition of multiple, functionally distinct TATA elements, one of which is a target in the hsp70 promoter for E1A regulation

https://doi.org/10.1016/0092-8674(88)90410-2Get rights and content

Abstract

We have dissected the human hsp70 promoter to define sequence elements allowing response to E1A. Alterations of sequence upstream of the TATA element, either with Bal 31 nuclease or by site-directed mutagenesis, had little or no effect on the response of the promoter to E1A. In general, the basal level was reduced, indicating that these sites interact with factors important for transcription, but regulation persisted. Although a CAT gene driven by just the hsp70 TATA (void of upstream sequences) could be stimulated by E1A, a similar construct containing the early SV40 TATA element was not. Analysis of several additional such constructions indicated that the specific sequence TATAA was crucial. Substitution of the TATAA sequence with the SV40 TATTAA element in the context of the wild-type hsp70 promoter resulted in loss of E1A inducibility, but maintenance of heat inducibility. Replacement of this element with sequences not related to any TATA element resulted in loss of activity and inducibility. Thus, the SV40 TATA equivalent is functional in the context of the hsp70 promoter but cannot be induced by E1A. We conclude that the target for E1A induction of the hsp70 promoter is TATAA, and that multiple functionally distinct TATA elements, and presumably cognate transcription factors, can be distinguished in eukaryotic cells.

References (47)

  • H.R.B. Pelham

    A regulatory upstream promoter element in the Drosophila hsp70 heat shock gene

    Cell

    (1982)
  • C. Benoist et al.

    In vivo sequence requirements of the SV40 early promoter region

    Nature

    (1981)
  • A.J. Berk et al.

    Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease digested hybrids

    Cell

    (1977)
  • E.Y. Chen et al.

    Supercoil sequencing: a fast and simple method for sequencing plasmid DNA

    DNA

    (1985)
  • C.M. Corsaro et al.

    Enhancing the efficiency of DNA-mediated gene transfer in mammalian cells

    Com. Cell Genet.

    (1981)
  • E.A. Craig

    The heat shock response

    Crit. Rev. Biochem.

    (1985)
  • B. Drabent et al.

    In vitro transcription of a human hsp70 heat shock gene by extracts prepared from heat-shocked and non-heat-shocked human cells

    Nucl. Acids Res.

    (1986)
  • M.J. Ensinger et al.

    Selection and preliminary characterization of temperature-sensitive mutations of type 5 adenovirus

    J. Virol.

    (1972)
  • T.M. Fisch et al.

    c-fos sequences necessary for basal expression and induction by epidermal growth factor, 12-0-tetradecanoyl phorbol-13-acetate, and the calcium ionophore

    Mol. Cell. Biol.

    (1987)
  • P.K. Ghosh et al.

    Identification of a promoter component involved in positioning the 5′ terminus of simian virus 40 early mRNAs

  • P. Gilardi et al.

    The E4 transcriptional unit of Ad2: far upstream sequences are required for its transactivation by E1A

    Nucl. Acids Res.

    (1984)
  • P. Gilardi et al.

    The E4 promoter of adenovirus type 2 contains an E1A dependent cis-acting element

    Nucl. Acids Res.

    (1986)
  • C.M. Gorman et al.

    Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells

    Mol. Cell. Biol.

    (1982)
  • Cited by (208)

    • Multiple domains in the 50 kDa form of E4F1 regulate promoter-specific repression and E1A trans-activation

      2020, Gene
      Citation Excerpt :

      Perhaps p50E4F1 provides a remedy to such effects that would otherwise reduce PIC nucleation and/or re-initiation rates and thus diminish E1A-induced high-level E4 gene expression. It is noteworthy that in some contexts, substitution of non-canonical TATA box sequences in several other promoters also reduces their trans-activation by E1A (Nicholas and Nevins, 1991; Simon et al., 1988; Simon et al., 1990). In addition to TATA box sequences, a second notable difference between the E4 and pG5 promoters are binding sites for TFIIB, a TBP-interacting general transcription factor that plays an integral role in RNA polymerase II recruitment into the PIC and polymerase release from the complex to permit transcript elongation (Deng and Roberts, 2007; Juven-Gershon and Kadonaga, 2010; Thomas and Chiang, 2006).

    • Core promoter-specific gene regulation: TATA box selectivity and Initiator-dependent bi-directionality of serum response factor-activated transcription

      2016, Biochimica et Biophysica Acta - Gene Regulatory Mechanisms
      Citation Excerpt :

      However, enhancers and activators can also differentially stimulate canonical TFIID-dependent transcription in a core promoter-dependent manner. Early observations showed that different TATA box sequences have different abilities to convey the activating signals of certain enhancers and activators in mammalian cells [34,35] and in yeast [36–38]. The activation domains of VP16 and SP1 fused to the yeast GAL4 DNA-binding domain were also reported to have different core promoter preferences in mammalian cells, although no strict core promoter selectivity was observed since both VP16 and SP1 could significantly activate all core promoter types tested [39].

    • New problems in RNA polymerase II transcription initiation: Matching the diversity of core promoters with a variety of promoter recognition factors

      2007, Journal of Biological Chemistry
      Citation Excerpt :

      Analysis of molecular interactions between activator proteins and core promoter context-dependent PICs may argue for a model in which the core promoter is involved in processing information received on the cis-regulatory level of a gene. For example, different TATA box-containing promoters respond differentially to activators, suggesting that they do not bind the same TFIID and/or PIC complexes (65, 66). c-Fos and Elf-1 differentially activate transcription from TATA- and INR-containing core promoters (67, 68), whereas optimal induction by papilloma virus E2 and the artificial Gal-VP16 activator requires both TATA and INR elements (69).

    • Non-optimal TATA elements exhibit diverse mechanistic consequences

      2006, Journal of Biological Chemistry
      Citation Excerpt :

      Taken together, these studies suggest that the TBP-DNA complex may exist in different structural contexts for variant sequences, and this context may also impact downstream steps in the transcription process. Because variant TATA elements play a fundamental role in the proper expression of many eukaryotic genes (9, 10, 21, 24), it is essential to determine the mechanisms underlying differential levels of transcription from varying TATA elements. To further explore the role of sequence variation in TATA element function, the in vivo and in vitro properties of a panel of substituted elements were characterized (summarized in Table 3).

    View all citing articles on Scopus
    §

    Present address: Howard Hughes Medical Institute, Department of Microbiology-Immunology, Duke University Medical Center, P. O. Box 3054 Durham, North Carolina 27710.

    View full text