Gene Regulation, Part 1 Lecture 15 Fall 2008 1 Metabolic Control in Bacteria • Regulate enzymes already present – Feedback Inhibition – Fast response • Control production of enzymes – Regulates transcription – Longer-term response Fig. 18.2 2 Metabolic Control in Bacteria • Operon – Unit of genetic function, found in bacteria and phages, consisting of an operator, promoter & coordinately regulated cluster of genes whose products function in a common pathway 3 Metabolic Control in Bacteria • Promoter – Specific area of DNA that designates the start of a gene – Where RNA polymerase binds • Operator – Sequence of nucleotides near the start of an operon to which an active repressor can attach – On-off “switch” for the cluster of genes Fig. 18.3 4 Metabolic Control in Bacteria • Regulatory gene – Gene that codes for a protein that controls the transcription of another gene or group of genes • Repressor – Protein that inhibits gene transcription – Binds to operator & prevents RNA polymerase from attaching to promoter Fig. 18.3 Metabolic Control in Bacteria • Repressors – Specific to an operator of a specific gene • Regulatory genes expressed continuously, but at low rate – On-off regulated by concentration of repressors – Repressors are allosteric proteins • Active & inactive form • Corepressors – Small molecules that cooperate with repressor to turn off transcription 5 6 Metabolic Control in Bacteria • Repressible Operon – Operon where transcription is normally “on”, but that can be inhibited (repressed) – E.g., trp operon (tryptophan) Fig. 18.3 Metabolic Control in Bacteria trp operon • trp repressor synthesized in an inactive form • Tryptophan acts as corepressor Fig. 18.3 7 8 Metabolic Control in Bacteria • Inducible operon – Operon where transcription is normally “off”, but that can be stimulated (induced) – E.g., lac operon (lactose) • lac repressor synthesized in active form • Binds to operator • Prevents RNA Polymerase from binding Fig. 18.4 Metabolic Control in Bacteria • Inducer binds to lac repressor • Inducer: specific small molecule that binds to repressor protein and changes its shape – Repressor inactivated – Releases/does not bind with operator – Transcription occurs Fig. 18.4 9 Negative and positive gene regulation • Negative gene regulation – Operons switched off by active form of a repressor protein – E.g., lac and trp operons • Positive gene regulation – Regulatory protein interacts directly with the genome to switch transcript on 10 11 Negative and positive gene regulation Lactose metabolism only occurs when glucose concentrations low • Catabolite activator protein (CAP) & cyclic AMP (cAMP) • CAP is activator – Protein that binds to DNA and stimulates transcription of a gene • cAMP concentration high when glucose concentration low • cAMP activates CAP Fig. 18.5 Negative and positive gene regulation • CAP attaches to CAPbinding site on promoter • Increases affinity of RNA polymerase for promoter • Activators increase the rate of transcription – As long as lactose is present, transcription can still occur Fig. 18.5 12