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Most is known about initiation. A number of DNA regions (generally located upstream
from the initiation site) and protein factors that bind to these sequences to regulate the
initiation of transcription have been identified.
• The processes of DNA and RNA synthesis are similar in that they involve
(1) the general steps of initiation, elongation, and termination with 5' to 3' polarity;
(2) large, multicomponent initiation complexes; and
(3) adherence to Watson-Crick base-pairing rules.
The RNA transcript with a 5' to 3' polarity is complementary to the template strand
with its 3‘ to 5' polarity. Note that the sequence in the RNA transcript and its polarity
is the same as that in the coding strand, except that the U of the transcript replaces
the T of the gene.
The Template Strand of DNA Is Transcribed
• The strand that is transcribed or copied into an RNA molecule is referred to as the
template strand of the DNA. The other DNA strand, the non-template strand, is
frequently referred to as the coding strand of that gene. It is called this because,
with the exception of T for U changes, it corresponds exactly to the sequence of the
RNA primary transcript, which encodes the (protein) product of the gene.
• In the case of a double-stranded DNA molecule containing many genes, the template
strand for each gene will not necessarily be the same strand of the DNA double helix
Thus, a given strand of a double-stranded DNA molecule will serve as the template
strand for some genes and the coding strand of other genes.
Note that the template strand is always read in the 3' to 5' direction.
The Prokaryotic Transcription
• The transcription "bubble" is an approximately 20-bp area of melted DNA, and the
entire complex covers 30–75 bp, depending on the conformation of RNAP.
The transcription cycle in bacteria
Bacterial RNA transcription is described in five steps:
(1) Template binding: RNAP binds to DNA and locates a promoter (P) melts
the two DNA strands to form a preinitiation complex (PIC).
(4) Chain elongation: Successive residues are added to the 3'-OH terminus of
the nascent RNA molecule.
(5) Chain termination and release: The completed RNA chain and RNAP are
released from the template. The RNAP holoenzyme re-forms, finds a
promoter, and the cycle is repeated.
Bacterial promoters, E Coli
• The inverted repeat, when transcribed into RNA, can generate the hairpin secondary
structure in the RNA transcript. Formation of this RNA hairpin causes RNAP to pause
and subsequently the termination factor interacts with the paused polymerase and
somehow induces chain termination.
• Transcription continues into the AT region, and with the aid of the ρ termination
protein the RNA polymerase stops, dissociates from the DNA template, and releases
the nascent transcript.
Rho-dependent transcription termination signals
• The inverted repeat, when transcribed into RNA, can generate the hairpin secondary
structure in the RNA transcript. Formation of this RNA hairpin causes RNAP to pause
and subsequently the termination factor interacts with the paused polymerase and
somehow induces chain termination.
• Transcription continues into the AT region, and with the aid of the ρ termination
protein the RNA polymerase stops, dissociates from the DNA template, and releases
the nascent transcript.
Eukaryotic Promoters Are More Complex
Schematic diagram showing the transcription control regions in a hypothetical
mRNA-producing, eukaryotic gene transcribed by RNA polymerase II. Such a gene
can be divided into its coding and regulatory regions, as defined by the transcription
start site (arrow; +1).
Proximal and distal cis elements are bound by trans -acting transcription factors, in
this example: Sp1 and CTF (also called C/EBP, NF1, NFY). These cis elements can
function independently of orientation (arrows).
Basal Transcription Complex
• Formation of the basal transcription complex begins when TFIID binds to the TATA
box. It directs the assembly of several other components by protein-DNA and
protein-protein interactions; TFIIA, B, E,F, H, and polymerase II (pol II).
The entire complex spans DNA from position –30 to +30 relative to the initiation site
(+1, marked by bent arrow)
Promoter Accessibility
and Hence PIC
Formation
Is Often Modulated
by Nucleosomes
Nucleosome eviction by
chromatin-active coregulators
facilitates PIC formation and
transcription.
Transcription Factors
Two Models Can Explain the Assembly of the
Preinitiation Complex
• RNA MOLECULES ARE USUALLY PROCESSED BEFORE THEY BECOME
FUNCTIONAL
TFIID, TFIIA, TFIIB, TFIIE, TFIIH, TFIIF, Med Binding of a single protein complex:
pol II, Med and six GTFs
The central dogma of molecular biology. Solid arrows indicate the types of
genetic information transfers that occur in all cells. Special transfers are indicated by
the dashed arrows: RNA-directed RNA polymerase occurs both in certain RNA
viruses and in some plants (where it is of unknown function); RNA-directed DNA
polymerase (reverse transcriptase) occurs in other RNA viruses; and DNA directly
specifying a protein is unknown but does not seem beyond the realm of possibility.
However, the missing arrows are information transfers the central dogma postulates
never occur: protein specifying either DNA, RNA, or protein. In other
words, proteins can only be the recipients of genetic information.
Gene expression. One strand of DNA directs the synthesis of RNA, a process
known as transcription. The base sequence of the transcribed RNA is complementary
to that of the DNA strand. The RNAs known as messenger RNAs (mRNAs) are
translated when molecules of transfer RNA (tRNA) align with the mRNA via
complementary base pairing between 3-nucleotide segments known as codons.
Each type of tRNA carries a specific amino acid. These amino acids are covalently
joined by the ribosome to form a polypeptide. Thus, the sequence of bases in DNA
specifies the sequence of amino acids in a protein.
Function of the transcription bubble