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James 4:7 7 Submit yourselves therefore to God. Resist the devil, and he will flee from you.

James 4:7 7 Submit yourselves therefore to God. Resist the devil, and he will flee from you. Regulation of Transcription. Timothy G. Standish, Ph. D. Eukaryotic RNA Polymerase II Promoters.

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James 4:7 7 Submit yourselves therefore to God. Resist the devil, and he will flee from you.

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  1. James 4:7 7Submit yourselves therefore to God. Resist the devil, and he will flee from you.

  2. Regulation of Transcription Timothy G. Standish, Ph. D.

  3. Eukaryotic RNA Polymerase II Promoters • Eukaryotic promoters are made up of a number of sequence elements spread over about 200 bp upstream from the transcription start site • In addition to promoters, enhancers also influence the expression of genes • Control of gene expression in eukaryotes involves many more factors than control in prokaryotes • This allows much finer control of gene expression

  4. Exon 1 Promoter 5’ Sequence elements TATA ~200 bp Some sequence elements are also response elements Eukaryotic Promoters

  5. Response Elements • Response elements are short sequences found either within about 200 bp of the transcription start site, or as part of enhancers • Different genes have different response elements • Binding of transcription factors to response elements determines which genes will be expressed in any cell type under any set of conditions

  6. Heat Shock Response Elements • Sudden changes in the temperature of cells cause stress in response to which heat shock genes are expressed • At least some heat shock genes are thought to be chaperones that help proteins fold correctly • Heat shock genes have Heat Shock Elements (HSEs) in their control regions • Heat Shock Transcription Factors (HSTFs) bind the HSEs up regulating expression of heat shock gene products

  7. Hot Cool Heat shock genes HEAT! Heat shock response elements The Heat Shock Response HSTF Kinase

  8. Hot Cool Heat shock response elements The Heat Shock Response Activation of kinase HSTF Kinase Phosphorylation of HSTF

  9. Hot Cool Heat shock response elements The Heat Shock Response Activation of HSTF Kinase HSTF Transcription initiation of all heat shock genes with HSEs

  10. Metallothionein Regulation • Heat shock protein regulation illustrates how a group of genes can be regulated by a single transcription factor • Other genes are regulated by a group of transcription factors which allows them to either respond to multiple separate situations, or respond only to a specific combination of situations • Metallothionein is an example of a gene that can be turned on by multiple transcription factors

  11. TGACTCA A consensus sequence to which AP1 binds conferring a response to phorbol esters Glucocorticoid Response Element - Behaves as an enhancer and allows regulation by steroids GRE BLE MRE MRE BLE MRE GC MRE TATA TRE -260 -240 -220 -200 -180 -160 -140 -120 -100 -80 -60 -40 -20 Metallothionein Regulator Elements - Multiple copies confer greater levels of induction by heavy metals Basal Level Elements (needed for transcription, act as enhancers) Constitutive Elements Metallothionein Regulation

  12. Regulation By TFs • A single transcription factor (or group of transcription factors) may regulate expression of a group of genes (i.e., heat shock proteins) • A single gene may be regulated by a number of independent transcription factors (i.e., metallothionein) • Eukaryotic regulation does not seem to involve repression • To achieve high levels of expression, several different transcription factors binding to different response elements may be necessary

  13. How Do TFs Recognize Response Elements? • Some mechanism must exist for protein transcription factors to recognize the specific sequences of bases found in response elements • There is no known specific relationship between a given amino acid and a base that would allow direct recognition • Groups on bases that are not involved with base pairing have the potential to be recognized by proteins • Some specific sequences are known to influence the gross structure of DNA

  14. DNA Binding Domains • Transcription factors exhibit a number of different motifs found in the area known to bind DNA: • Zinc finger -First found in TFIIIA • Helix-turn-helix - First described from phage receptors • Amphipathic Helix-loop-helix - Identified in some development regulators • Leucine zipper - Held together by interactions between leucine amino acids

  15. Zinc Fingers • Are found in steroid receptors and are common in other transcription factors • Interaction between cysteine and histidine amino acids and the divalent zinc ion results in the formation of loops called “fingers” • One side of each loop forms an a-helix which can lie in the major grove of DNA • Interaction between the amino acids in the zinc finger and bases in the DNA allow for sequence recognition

  16. O O H2N C OH C H2N C H OH H C C H H H C HS H C H C N H N C Zinc Fingers

  17. ≈ 6 Amino acids ≈ 23 Amino acids C C C H H H 2- 4 Amino acids Zn++ Zn++ Zn++ C C H H C H 7 - 8 Amino acid linker Zinc Fingers

  18. The End

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