Heat shock promoter

Goldenbetg, C.J., Luo, Y., Fenna, M., Baler, R., Weinmann, R., Voellmy, R. (1988). Purified human factor activates heat shock promoter in a HeLa cell free transcription system. J. Biol. Chem. 263, 19734-19739. 

Lis, J. Wu, C. (1993). Protein traffic on the heat shock promoter-parking, stalling, and trucking along. Cell 74, 1-4. 

Tsukiyama, T., Becker, P.B., Wu, C. (1994). ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor. Namre 367, 525-532. 

Pelham, H.R.B. Bienz, M. (1982). A synthetic heat-shock promoter element confers heat-inducibility on the herpes simplex virus thymidine kinase gene. EM BO Journal, 1,1473-7. 

Tsukiyama, T., Becker, P.B., and Wu, C. (1994) ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor. Nature 367, 525-532. Laurent, B.C., Yang, X., and Carlson, M. (1992) An essential Saccharomyces cerevisiae gene homologous to SNF2 encodes a helicase-related protein in a new family. Mol. Cell Biol. 12, 1893-1902. 

FIGURE 28-3 Consensus sequence for promoters that regulate expression of the E. coli heat-shock genes. This system responds to temperature increases as well as some other environmental stresses, resulting in the induction of a set of proteins. Binding of RNA polymerase to heat-shock promoters is mediated by a specialized a subunit of the polymerase, cr32, which replaces cr70 in the RNA polymerase initiation complex. 

Induction of heat-shock proteins depends upon a heat-shock promoter element (HSE) that binds an activating transcription factor HSF.452-455 An increase in temperature not only induces synthesis of heat-shock proteins but represses synthesis of most other proteins. Thus, in E. coli or Salmonella a shift from 30°C to 42°C causes the appearance of 13 heat-shock proteins. At 50°C synthesis of almost all other proteins stops. In E. coli transcription of heat-shock genes is controlled by alternative factors, o32 and oE.456 456a... 

Functional analysis of heat shock promoter elements... 

Czarnecka, E., Key, J.L. Gurley, W.B. (1989). Regulatory domains of the Gmhspl7.5-E heat shock promoter of soybean. Molecular and Cellular Biology 9, 3457-63. 

Schoffi, F., Rieping, M., Baumann, G., Bevan, M.W. Angermiiller, S. (1989). The function of plant heat shock promoter elements in the regulated expression of chimaeric genes in transgenic tobacco. Molecular and General Genetics 217, 246-53. 

Heat inducible gene expression systems can be activated by a variety of technologies, all via the production of hyperthermia. Technologies used in clinical applications of hyperthermia include simple approaches, such as water baths, or more sophisticated methods, such as microwave or radiofrequency radiations and ultrasound (Gemer and Cetas, 1993). The magnitude of heat shock promoter activity induction is dependent on both the time of exposure to hyperthermia and the hyperthermic temperature (Gemer et al., 2000). HSP promoter activity is activated by temperature in a species-specific manner. In flies, HSP promoter activity is activated by temperatures over 30 °C (Lindquist, 1986). In human cells, temperatures of 40 °C and above are required to activate HSP promoters (Gemer et al., 2000). 

Standard promoter Z Heat-shock promoter 3 Nitrogen-starvation promoter... 

Figure 28.6. Alteruative Promoter Sequeuces. A comparison of the consensus sequences of standard promoters, heat-shock promoters, and nitrogen-starvation promoters of E. coli. These promoters are recognized hy a a 32 and a respectively.

Several copies of this sequence, known as the heat-shock response element, are present starting at a site 15 bp upstream of the TATA box. HSTF differs from a 32 a heat-shock protein of E. coli (Section 28.1.2). in binding directly to response elements in heat-shock promoters rather than first becoming associated with RNA polymerase. 

Figure 28.22. Transcription-Factor-Binding Sites. These multiple binding sites for transcription factors were mapped by footprinting. (A) Binding of Spl (green) to the SV40 viral promoter and to the dihydrofolate reductase (DHFR) promoter. (B) Binding of HSTF (blue) to a.Drosophila heat-shock promoter. [After W. S. Dynan and R. Tjian. Nature...

Kowalski, J. Gilbert, S.A. van Drunen-Littel-van den Hurk, S. van den Hurk, J. Babiuk, L.A. Zamb, T.J. Heat-shock promoter-driven synthesis of secreted bovine herpesvirus glycoproteins in transfected cells. Vaccine 1993, II, 1100-1107. 

Induction of heat-shock proteins depends upon a heat-shock promoter element (HSE) that binds an activating transcription factor An increase... 

Explain how the a factor enables RNA polymerase to recognize promoters. Distinguish between the a70 and a32 subunits, contrast the sequences of standard promoters and heat-shock promoters, and provide examples of how a factors can determine which genes are... 

A similar stepwise mechanism for transcriptional activation has also been proposed on the basis of studies of recruitment of the transcriptional activator HSF, Mediator, and RNA Pol II to the heat shock promoter of Drosophila polythene chromosomes (Park et al., 2001). Using different techniques, it was observed that on heat shock, both HSF and Mediator are rapidly recruited to the hsp70 promoter in a manner that is independent of the presence of a core promoter. This recruitment was not accompanied with a corresponding increase in RNA Pol II or GTFs and was also independent of the presence of the RNA Pol II inhibitor ct-amanitine. 

Bond, U. Schlesinger, M.J. (1986). The chicken ubiquitin gene contains a heat shock promoter and expresses an unstable mRNA in heat shock cells, h/lol Cell Biol, 6, 4602-10. 

The initial purification to homogeneity took 4 weeks of uninterrupted work of one investigator and an assistant, in addition to the recruited assistance of 3 members of the laboratory. Column fractions were assayed by Exonuclease III protection of an end-labeled heat shock promoter fragment. Current purifications are handled adequately by a research assistant, and each purification is completed in about two weeks this is aided by the use of the more convenient gel mobility shift assay. A sample of the protein content in the active fractions at various stages of the purification is shown in Figure 2. 

HSE is needed for recruitment of the RNA polymerase to the heat-shock promoter. The binding of TFIID to the uninduced promoter may help heat-shock genes respond more rapidly to an increase in temperature. 

The gene for Drm-SP-1 has been localized by in situ hybridization to polytene chromosomes at 70A (20). This 266 bp single-copy gene with one intron has been cloned and expressed when linked to the heat shock promoter or the yolk protein 1 promoter (E. Kubli and coworkers as cited by (41)). Other aspects of Drm-SP-1 molecular biology are summarized by Chen (41). 

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