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Molecular Basis for Relationship between Genotype and Phenotype genotype DNA DNA sequence transcription RNA translation protein function phenotype organism amino acid sequence Enhanceosomes and Synergistic Effect on Transcription Enhanceosome: protein complex of trans-acting factors bound to appropriate DNA sequences. Proteins interact synergistically to elevate transcription rate. In b-interferon gene transcription, TFs recruit a coactivator (CBP) which is needed for transcription to occur normally. Formation of the enhanceosome and activation of RNA polymerase by coactivator are necessary for efficient transcription. Transcription of b-interferon gene is activated during viral infection. b-interferon enhanceosome: • Surrounded by two nucleosomes, nuc1 and nuc2. • nuc2 covers TATA box and transcription start site. • Co-activator GCN5 acetylates nuc1 and nuc2. • Acetylation leads to recruitment of co-activators, chromatin remodeling complex, and RNA pol II. Tissue-specific Regulation of Transcription Regulated transcription depends on: - specific enhancer for gene(s) - enhancer-specific activator proteins - correct interaction between enhancer and activator Tissue-specific regulation requires that the enhancer-specific activator is present only in cells of that tissue type. ectopic expression: expression in an abnormal location “Master Switch” Gene Eye formation requires over 2000 genes. eyeless (ey) mutation causes small rudimentary eyes to form in Drosophila melanogaster. Small eyes (Sey, Pax-6) in mouse causes similar phenotype. Aniridia gene in human (lack of normal iris) shows considerable homology to ey gene. Comparison of ey+ and ey Phenotypes Wild-type eyes eyeless (ey) eyes size of ey eyes “Master Switch” Gene Wild-type eyeless (ey) gene can be induced to be expressed ectopically. eyeless (ey) gene codes for a helix-turn-helix transcription protein.