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3) Gene Control in Bacteria - Transcription Activators and Repressors You are interested in examining the regulation of the gene that encodes an enzyme, Tre- ase, important in metabolizing trehalose into glucose in bacteria. Trehalose is a disaccharide formed of two glucose units. It is known that two DNA-binding proteins, TreA and TreB, are important for binding to the promoter of the Tre-ase gene and are involved in regulating the transcription of the Tre-ase gene: TreA binds to the A site in the promoter region, and TreB binds to the B site. You make mutations in the TreA and TreB genes to create cells lacking these genes, observe what happens to transcription of the Tre-ase gene,and obtain the results in the table below: Transcription of Tre-ase in different media Glucose Trehalose normal cells lacking TreA cells lacking TreB cells lacking both TreA &TreB on OFF OFF OFF OFF on ON ON OFF OFF Glucose + Trehalose OFF ON OFF OFF a. What is the role for TreA in controlling Tre-ase expression? Explain. b. What is the role for TreB in controlling Tre-ase expression? Explain. c. From these data, what do you predict will happen to Tre-ase transcription (compared with that in normal cells) in the presence of trehalose if you were to create a version of the TreA protein that will constitutively bind to the A site in the Tre-ase promoter?

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Ans a) Tre A is an activator protein. When it is present, in the absence of glucose, cells actively transcribe the trehalose operon (trehalose only, normal cells). However, when environmental conditions are the same but Tre A is absent (trehalose only, lacking Tre A), transcription continues [because the repressor is unbound] but only at a low level—cells cannot be activated to transcribe the operon at a high level.

Ans b)Tre B is a repressor protein that, when absent, results in cells that have constitutive expression (i.e., “always on”) of the Trehalose operon , even in the absence of trehalose (glucose only).

Ans c) Since the data presented support that Tre B is a repressor protein, if a version of the protein were always bound to the A element of the promoter, transcription of the trehalose operon will be able to proceed—even in the presence of the carbohydrate.

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