Eukaryotic Regulation

Eukaryotic Regulation - Lecture 7: DNA binding proteins and...

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Lecture 7: DNA binding proteins and Gene Expression in Eukaryotes Reading: Chapter 11 385-389; 394- 397; 402-411
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Comparison of Regulation of Gene Expression Between Prokaryotes and Eukaryotes Control of transcription through specific DNA-binding proteins Role played by chromatin structure Coordination achieved by operons Differential splicing Attenuation Differential polyadenylation Differential transport from nucleus to cytoplasm Differential rates of translation Prokaryotes Eukaryotes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes No No No No No No
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Similar in design to prokaryotes, but in general more complex. The rate of transcription of a gene is the sum of positive and negative regulatory influences exerted by proteins bound to cis acting elements. • The level of transcription is controlled by regulating the rate of initiation. Transcriptional Regulation in Eukaryotes
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Cis-Acting sequences
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Core Promoter in Eukaryotes TATA Box Promoter proximal elements Core Promoter Core promoter alone NOT able to promote transcription - requires activating elements
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Can act up more than 50 kb away (though they can also be close) Can be either upstream or downstream of the coding region Can act in either orientation. Can either elevate the rate of transcript initiation ( enhancers ) or reduce the rate of initiation ( silencers ) Distant-independent elements
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core promoter promoter-proximal elements transcript scale is in kb (kilobases) enhancers +1 Different enhancers of dpp promoter
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Reporter Genes Allow the Analysis of Promoters lacZ gene Core promoter Your favorite enhancer Transform into organism Analyze pattern of reporter activity A reporter is a gene that encodes a product that is easy to assay and gives an insoluble product
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Deletion analysis of promoters – Identification of cis acting sites Core promoter lacZ gene Your favorite promoter lacZ gene lacZ gene lacZ gene lacZ gene Lac Z activity lacZ gene lacZ gene lacZ gene lacZ gene + - - - + + + - + lacZ gene +
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Transgenic Drosophila harboring a fusion of the dpp promoter to lacZ LacZ in situ imaginal disk LE enhancer Different enhancers of dpp promoter Coding region Kb (1000 bps)
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GFP reporter showing expression of an odorant receptor gene in C. elegans GFP reporter in adult mice GFP as a Reporter Gene
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Trans-Acting Factors
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: Trans acting factors that alter the rate of transcription. Often bind to DNA. Transcriptional activators : increase rate of transcription. Transcriptional repressors
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Eukaryotic Regulation - Lecture 7: DNA binding proteins and...

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