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CONTROL MECHANISMS Sections 5.5 Page 255 Outline 1. 2. 3. 4. 5. Gene regulation Operons lac operon trp operon Recap Gene regulation Housekeeping genes - Always needed; constantly being transcribed/translated Most genes are not housekeeping genes Gene regulation involves turning certain genes on or off ◦ depends on whether their products are required Regulators are the proteins that switch genes on or off ◦ activators, or ◦ repressors Recall… Protein synthesis occurs in two distinct stages: 1. Transcription, in the _________ ◦Product: ____ 2. Translation, in the ______________ ◦Product: __________ Controlling gene expression Levels of control 1. Transcriptional 2. Posttranscriptional 3. Translational 4. Posttranslational Levels (cont’d) Transcriptional gene regulation Regulation of which genes are transcribed Can involve control of the rate at which transcription occurs Posttranscriptional gene regulation Modifications made to primary transcript in the nucleus Take a break… Can you make an educated guess about: Translational control?? Post-translational?? Levels (cont’d) Translational gene regulation Control of how often and how rapidly mRNA transcripts are translated into proteins Posttranslational gene regulation Modifications made to the polypeptide chain Transcriptional regulation Two classic examples: lac operon – negative regulation trp operon – positive regulation What is an operon? only in prokaryotic cells a cluster of genes all genes are under the control of one set of regulatory sequences ◦ promoter and operator sequences ◦ regulation is dependent on the presence or absence of effector molecules Operator sequence Sequence to which regulator protein binds Depending on the regulator, binding to the operator causes either: ◦ activation of transcription ◦ repression of transcription Effector Acts on the regulator protein Can be an inducer ◦ stimulates transcription Can be a co-repressor ◦ inhibits transcription THE LAC OPERON Lac operon lactose = glucose + galactose disaccharide found in milk E. coli: ß-galactosidase catalyses the cleavage of the bond in lactose Not economical to produce ß-galactosidase when lactose is not present Lac operon (cont’d) Negative control mechanism prevents production of ß-galactosidase if lactose is absent Structure of lac operon Cluster of genes: lacZ, lacy, lacA Each codes a different part of the enzyme ALL are under the control of one promoter LacI protein – repressor protein. Blocks transcription of the lac operon by binding to the operator. Physical blockage: Covers part of the lac promoter Mechanism of regulation http://www.sumanasinc.com/webcontent/ animations/content/lacoperon.html Mechanism of regulation Mechanism of regulation lactose is absent •LacI binds to lac promoter. •No transcription. lactose is present Lactose binds to LacI protein. LacI changes conformation → unable to bind lac operator. RNA polymerase can access lac promoter; transcription occurs. What is the effector molecule? _______________ It acts as an inducer – it induces transcription by deactivating the repressor protein. Negative control Default: Transcription is off, due to binding of a repressor protein. Presence of an effector (inducer) removes the repressor from the operator region. Transcription can now occur. TRP OPERON The trp operon Tryptophan – Amino acid Operon: cluster of five genes encoding enzymes required for trp synthesis Transcription of operon is repressed when concentrations of trp in the cell are high trp operon (cont’d) Positive control mechanism prevents production of tryptophan, if tryptophan levels are high http://highered.mcgrawhill.com/olc/dl/120080/bio26.swf Mechanism of regulation Mechanism of regulation trp absent •trp operon is transcribed (default) trp present •trp itself will bind to the repressor protein. •conformational shape occurs •allows repressor to bind to the trp operator → shuts off transcription What is the effector molecule? _______________ It acts as a co-repressor. It binds to a repressor to activate it in order to repress transcription. Positive control Default: Transcription is on Presence of an effector (co-repressor) causes binding of the repressor to the operator. Transcription does not occur. trp vs. lac operons Default state Positive control Effector Transcription is on Allows repressor to bind Negative Transcription control is off Removes repressor Positive vs. Negative Control: Is the repressor added or taken away? Recap: Vocabulary Basic terms: Gene regulation: Describes any process that alters the rate of gene expression. Operon: A cluster of closely-related genes. They are all controlled by one set of regulatory sequences. Types of molecules involved: Regulators: Molecules that carry out translational gene regulation. ◦ Classified as either activators, or repressors. ◦ Example: LacI repressor; trp repressor Effectors: Substances to which regulator proteins respond. ◦ Inducers or Co-repressors ◦ Example: Lactose, Tryptophan Regulatory DNA sequences: Promoter: DNA sequence to which RNA polymerase binds to begin transcription. Operator: DNA sequence to which a transcription factor binds in order to alter transcription. Close to the promoter. Homework Pg. 258 #1-6