Transcription
An
important feature of RNA synthesis is that even though the DNA molecule being
copied is double stranded, in any particular region of DNA only one strand
serves as a template. The DNA strand copied into RNA molecule is called CODING
OR SENSE STRAND.
The
synthesis of RNA consists of five discrete stage (Figure 12.2):
1. Promoter recognition:
RNA polymerase binds to DNA within a specific base sequence (20–200 bases long)
called a promoter. The sequence TATAAT (or a nearly identical sequence) often
called a pribnow box or – 10 region is found as part of all prokaryotic
promoters.
The RNA
polymerase of the bacterium E.coli
consists of five protein subunits. Four of the subunits comprise the core
enzyme (catalyzes the joining of the nucleoside triphosphates to the RNA) and
fifth subunit, the σ subunit (required for promoter binding).
2. Local unwinding of DNA occurs and RNA polymerase forms an open promoter complex.
3. The first nucleoside triphosphate is placed at
polymerization start site (near to the initial binding site) and synthesis
begins.
4. RNA polymerase then moves along the DNA, adding
ribonucleotides, to the growing RNA chain.
5. RNA polymerase reaches chain termination
sequence and both the newly synthesized RNA and the polymerase are released.
Two kinds of termination events are known those that are self – terminating
(dependent on the base sequence only) and those that require the presence of
the termination protein Rho.
Initiation
of a second round of transcription need not await completion of the first, for
the promoter becomes available once RNA polymerase has polymerized 50–60
nucleotides. In bacteria most mRNA molecules are degraded within a few minutes
after synthesis. This degradation enables cells to dispense with molecules that
are no longer needed.
In
prokaryotes mRNA molecules commonly contain information for the amino acid
sequences of several different polypeptide chains. In this case, such a
molecule is called polycistronic mRNA. Cistron is a term used to mean a base
sequence encoding a single polypeptide chain. The genes contained in
polycistronic mRNA molecule (Figure 12.3) often encode the different portions
of a metabolic pathway. For example, in E.
coli the ten enzymes needed to synthesize histidine are encoded in one mRNA
molecule .
In prokaryotes the immediate product of transcription (called the primary transcript) is mRNA, in contrast in eukaryotes the primary transcript must be converted to mRNA. This conversion called RNA processing consists of two types of events- modification of termini and excision of untranslated sequences (non-coding sequence or introns) embedded within coding sequences (exons). Introns excision and the joining of exons to form an mRNA molecule is called RNA splicing. The introns are present in almost all eukaryotic transcripts but are rare in the free – living unicellular eukaryotes such as yeast. Some bacterial genes do contain introns.
Ribosomal
RNA and tRNA are also transcribed from genes. The production of these molecules
is not as direct as synthesis of bacterial mRNA. The main difference is that
these RNA molecules are excised from large primary transcripts. Highly specific
RNA excise rRNA and tRNA from these large transcripts, and other enzymes
produce the modified bases in tRNA.
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