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8.21:

Transcription Initiation

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Molecular Biology
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JoVE Core Molecular Biology
Transcription Initiation

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For RNA Polymerase II, the most well-studied promoter is called the TATA box. It has a conserved DNA sequence, most commonly T-A-T-A-A-A, which is usually located 25 nucleotides upstream from the transcription start site. RNA polymerase II is guided to the promoter site by a set of proteins known as the general transcription factors, specifically Transcription Factor 2, or TFII, with variants A, B, D, E, F, and H. Transcription begins with the binding of TFIID, to the TATA box. The TATA box Binding Protein, or TBP, which is a component of TFIID, recognizes the TATA box DNA sequence. Next, TBP associates with TFIIA and TFIIB, building a platform for RNA Polymerase to assemble with TFIIF at the promoter site. Finally, TFIIE and TFIIH join these components to form the initiation complex. Next, TFIIH unwinds the DNA duplex around the start site and phosphorylates the C-terminal domain of the RNA polymerase. This phosphorylation changes the conformation of the polymerase, allowing it to release from the initiation complex and begin transcription at the start site. Once RNA polymerase II has started synthesizing the RNA transcript, most of the general transcription factors are released from the DNA.

8.21:

Transcription Initiation

Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.

The promoters and enhancers and their accessory proteins allow tight regulation of transcription. The TATA box is the most widely studied among the different core promoter sequences, though it is only present in 10-15% of mammalian genes. The TATA box provides a platform for assembling the various transcription factors (TF), including TFIID, which contains the TATA-binding protein subunit. This assembly of transcription factors with the RNAP forms the pre-initiation complex (PIC). The PIC alone has a low transcription rate that other proteins called activators and repressors can enhance or decrease.

Once the RNAP has begun elongation, the transcription factors are released from the DNA to initiate another round of transcription with a new RNA polymerase molecule. The RNAP binds strongly to the DNA template and synthesizes the RNA transcript for long distances without dissociating from DNA.

Suggested Reading

  1. Minakhin, Leonid, Sechal Bhagat, Adrian Brunning, Elizabeth A. Campbell, Seth A. Darst, Richard H. Ebright, and Konstantin Severinov. "Bacterial RNA polymerase subunit ω and eukaryotic RNA polymerase subunit RPB6 are sequence, structural, and functional homologs and promote RNA polymerase assembly." Proceedings of the National Academy of Sciences 98, no. 3 (2001): 892-897.
  2. Cooper GM. “The Cell: A Molecular Approach.” 2nd edition. Sunderland (MA): Sinauer Associates; (2000). Eukaryotic RNA Polymerases and General Transcription Factors.
  3. Kuehner, Jason N., Erika L. Pearson, and Claire Moore. "Unravelling the means to an end: RNA polymerase II transcription termination." Nature reviews Molecular cell biology 12, no. 5 (2011): 283-294.