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Controls Over Genes More on Transcription Promoters are regions on DNA that show where RNA Polymerase must bind to begin the Transcription of RNA Called the TATA box Transcription factors are also involved (proteins that mediate the binding of RNA polymerase) Specific base sequences act as signals to stop Called the termination signal mRNA Processing After the DNA is transcribed into RNA, editing must be done to the nucleotide chain to make the RNA functional Introns, non-functional segments of DNA are snipped out of the chain (RNA splicing) mRNA Editing Exons, segments of DNA that code for proteins, are then rejoined by the enzyme ligase A guanine triphosphate cap is added to the 5” end of the newly copied mRNA A poly A tail is added to the 3’ end of the RNA The newly processed mRNA can then leave the nucleus Result of Transcription CAP New Transcript Tail mRNA Transcript •mRNA leaves the nucleus through its pores and goes to the ribosomes Why Control Gene Expression? Some genes are “on” (being transcribed) almost all the time Called housekeeping genes Examples: ribosome components, enzyme for basic metabolic pathways Many genes are only turned on when they are needed Why Control? Transcribing genes that are not needed is a waste of energy and may interfere with the status of the cell Regulation Respond to a range of stimuli Prokaryotes respond to external stimuli (food, enzymes turned on) Eukaryotes also respond to internal stimuli (hormones, growth factors) Regulation Developmentally regulated Multicellular organisms progress through developmental stages Different genes expressed at different times during development Cell specialization Different genes expressed in different cells The strategy behind regulation.. Gene control is control over amount of gene produced (RNA or protein) in cell Multiple ways to control the amount of gene product in a cell Controlling gene product amount 1. Rate of transcription – rate mRNA is produced; faster produced = more product 2. mRNA degradation – rate mRNA is broken down; faster broken down = less product Controlling gene product amount 3. mRNA processing – capping, splicing; slower processing = less product 4. Translation – rate of translation or # of ribosomes translating; fast/more = more product Although control probably involves all of these, the most understood are changes in the rate of transcription Gene Control – lac operon Lac operon is a gene in bacteria Bacteria have 3 genes in a row (operon) that involve breaking down lactose for energy In order to be efficient, these genes should not be expressed unless lactose is present Lac Operon - vocab Regulatory protein – control transcription, translation, and gene products by interacting with DNA, RNA, or proteins Repressor – protein that binds with an operator on prokaryotic DNA to prevent transcription Operator – short base sequence between a promoter and genes; a binding site for repressors Lac operon – vocab. Promoter – piece of DNA where RNA polymerase can bind and start transcription Negative control – regulatory protein that slows down gene activity Positive control – regulatory protein that enhances gene activity Lac operon vocab. Operon – a promoter and a pair of operators that control a bacterial gene Activator – protein that exerts positive control over an operon Figure 15.3a Page 241 operator regulatory gene transcription, translation operator gene 1 gene 2 promoter lactose operon repressor protein gene 3 Lac operon Goal 1 – transcription low when lactose is absent Lac I (gene upstream from operon) produces a repressor which binds to promoter region Binding of repressor prevents RNA polymerase from binding and transcribing genes Lac operon Goal 2 – increase transcription when lactose is present Allolactose will bind to the repressor, changing its conformation and causing it to fall off the promoter site Promoter site now available for RNA polymerase to bind; transcription of lac genes begins Lac operon Goal 3 – turn off transcription when lactose is used up Allolactose metabolizes, freeing up the repressor The free repressor is available to bind the promoter site and stop transcription Control of lac operon Negative control – glucose present repressor inactivates the lac operon Positive control – lactose present – activator protein (called CAP) makes promoter more favorable for RNA polymerase to bind and begin transcription Low Lactose Repressor binds to operator Binding blocks promoter Transcription is blocked Figure 15.3b Page 241 High Lactose allolactose lactose mRNA operator promoter operator RNA polymerase gene 1 Figure 15.3c Page 241 Most Genes Are Turned Off Cells of a multicelled organism rarely use more than 5-10 percent of their genes at any given time The remaining genes are selectively expressed Homeotic Genes Occur in all eukaryotes Master genes that control development of body parts Encode homeodomains (regulatory proteins) Homeobox sequence can bind to promoters and enhancers X Chromosome Inactivation In female mammals, in all cells one of the two X chromosomes is completely inactivated Inactivation is random Inactivated chromosome can be observed in the interphase nucleus as Barr body Genes on the inactivated chromosome are not expressed