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Molecular Biology Notes
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Transcription is the first step in gene expression. It involves copying a gene's DNA sequence
to make an RNA molecule.
Transcription describes the process by which the genetic information contained within DNA
is re-written into messenger RNA (mRNA) by RNA polymerase. This mRNA then exits the
nucleus, where it provides the basis for the translation of DNA. By controlling the production
of mRNA in the nucleus, the cell regulates the rate of gene expression.
Both prokaryotes and eukaryotes perform fundamentally the same process of transcription,
with the important difference of the membrane-bound nucleus in eukaryotes. With the genes
bound in the nucleus, transcription occurs in the nucleus of the cell and the mRNA transcript
must be transported to the cytoplasm. In prokaryotes, which lack membrane-bound nuclei
and other organelles, transcription occurs in the cytoplasm of the cell. Thus, the processes
of transcription, translation, and mRNA degradation can all occur simultaneously.
RNA POLYMERASE
RNA Polymerase is the enzyme that produces the mRNA.
Prokaryotes use the same RNA polymerase to transcribe all of their genes. In E. coli, the
polymerase (holoenzyme) is composed of five polypeptide subunits. Four of these subunits,
denoted α, α, β, and β' comprise the polymerase core enzyme. These subunits assemble
every time a gene is transcribed, and they disassemble once transcription is complete.
Each subunit has a unique role; the two α-subunits are necessary to assemble the
polymerase on the DNA; the β-subunit binds to the ribonucleoside triphosphate that will
become part of the nascent “recently born” mRNA molecule; and the β' binds the DNA
template strand. The fifth subunit, σ, is involved only in transcription initiation.
Eukaryotic cells contain three distinct
nuclear RNA polymerases that
transcribe different classes of genes.
Stages of Transcription
The process of transcription can be broadly categorised into 3 main stages: initiation,
elongation & termination.
Initiation
Transcription in prokaryotes (and in eukaryotes) requires the DNA double helix to partially
unwind (with the help of helicase enzyme) in the region of mRNA synthesis. The region of
unwinding is called a transcription bubble. The DNA sequence onto which the proteins and
enzymes involved in transcription bind to initiate the process is called a promoter.
To begin transcription, the RNA polymerase holoenzyme recognises and assembles at the
promoter. The dissociation of σ allows the core enzyme to proceed along the DNA template,
synthesizing mRNA by adding RNA nucleotides according to the base pairing rules, similar
to the way a new DNA molecule is produced during DNA replication. Only one of the two
DNA strands is transcribed. The transcribed strand of DNA is called the template strand
because it is the template for mRNA production. The mRNA product is complementary to the
template strand and is almost identical to the other DNA strand, called the non-template
strand, with the exception that RNA contains a uracil (U) in place of the thymine (T) found in
DNA.
Elongation
Like DNA polymerase, RNA polymerase adds new nucleotides onto the 3′-OH group of the
previous nucleotide. This means that the growing mRNA strand is being synthesized in the
5′ to 3′ direction.
As elongation proceeds, the DNA is continuously unwound ahead of the core enzyme as the
hydrogen bonds that connect the complementary base pairs in the DNA double helix are
broken. The DNA is rewound behind the core enzyme as the hydrogen bonds are reformed.
The base pairing between DNA and RNA is not stable enough to maintain the stability of the
mRNA synthesis components. Instead, the RNA polymerase acts as a stable linker between
the DNA template and the newly forming RNA strand to ensure that elongation is not
interrupted prematurely.
Termination
Once a gene is transcribed, the prokaryotic polymerase needs to be instructed to dissociate
from the DNA template and liberate the newly made mRNA. Sequences called terminators
signal that the RNA transcript is complete.
Post-Transcriptional RNA Processing

DNA transcription occurs in a cell's nucleus. The RNA that is synthesized in this process is
then transferred to the cell's cytoplasm where it is translated into a protein. In prokaryotes,
the RNA that is synthesized during DNA transcription is ready for translation into a protein.
Eukaryotic RNA from DNA transcription, however, is not immediately ready for translation.
Post-transcriptional modifications of RNA accomplish two things: 1) Modifications help the
RNA molecule to be recognized by molecules that mediate RNA translation into proteins; 2)
During post-transcriptional processing, portions of the RNA chain that are not supposed to
be translated into proteins are cut out of the sequence. In this way, post-transcriptional
processing helps increase the efficiency of protein synthesis by allowing only specific protein-
coding RNA to go on to be translated. Without post-transcriptional processing, protein
synthesis could be significantly slowed, since it would take longer for translation machinery
to recognize RNA molecules and significantly more RNA would have to be unnecessarily
translated to achieve the same results.
Modification of DNA take place in three steps:

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Molecular Biology Notes

Post-Transcriptional RNA Processing

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