6-9. Passage Transcription Factors at An experiment was designed to determine th
ID: 89615 • Letter: 6
Question
6-9. Passage Transcription Factors at An experiment was designed to determine the sequental order in which transcription factors add to the initiation In different combinations, the transcription factors were added to radiolabeled (hot) DNA containing the promoter sequence and promoter proximal sequences, followed by on a native gel (no denaturation) and autoradiography. Seven different complexes of DNA and protein were observed (numbered 1-7 on the left side of the figure). aro formod when the same experiments are performed in the presence of excess None of the complexes amounts of unlabeled (cold) DNA sequences that include a TATAAA sequence. Complexes 2-7 still form in the presence of cold DNA sequences when the cold DNA TATAAA sequence AGAGA. Complex still does not form when the cold DNA sequence is AGAGAA. for the oompetition experiments with cold DNA are not shown, TFI IA. FIIB TFIID TFEIE RNA Pol II complex 7 free probe 2 3 4 5 6 7 8 9 10 11 12 6. What is the correct order of transcriptional events that can occur when RNA polymerase binds to the A Start of RNA synthesis, open complex formation, closed complex formation, promoter clearance B Closed complex formation, open complex formation, promoter clearance, start of RNA synthesis C Closed complex formation, open complex formation, start of RNA synthesis, promoter clearance D open complex formation, closed complex formation, start of RNA synthesis, promoter clearance E Promoter clearance, blue light special, closeout sale 7. Which one of the following best names the technique used in this experiment? A Westem Blot B DNA Footprinting C Electrophoretic Mobility Shift Assay (EMSA) D Sanger Sequencing E Northern BlotExplanation / Answer
Answer:
6. (C) Closed complex formation, open complex formation, start of RNA synthesis, promoter clearance
7. (C) Electrophoretic mobility shift assay (EMSA)
8. (C) TFIID
The complex 2 is actually the TFIID bound to the DNA. The increase in molecular weight is due to the presence of TFIIA.
TFIIA interacts with the TBP subunit of TFIID and aids in the binding of TBP to TATA-box containing promoter DNA. Although TFIIA does not recognize DNA itself, its interactions with TBP allow it to stabilize and facilitate formation of the Preinitiation complex.
9. (E) Some of the proteins in tha transcriptional apparatus do not recognize and/or bind to the promoter sequence.
Example: TFIIA interacts with the TBP subunit of TFIID and aids in the binding of TBP to TATA-box containing promoter DNA.
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