Transcription

What is transcription?
Transcription is the process where a molecule
of messenger RNA (mRNA) is synthesized
along a template strand of DNA.
Where does it happen?
DNA is actively transcribed in a region of the
nucleus called the nucleolus.
$teps in transcription
1) !art I-Initiation
!romoter is a sequence of DNA nucleotides
(15-300bp) that serve as the site where RNA
polymerase will recognize and bind.
n eukaryotes TATA box is typically present in
the promoter consisting of nucleotide
sequence like TATAAA.
TATA box aids in the recognition of promoter
First a group of proteins called transcription
factors locate and bind to the TATA box.
$tructure of the human TB! core domain compIexed
with DNA as determined by x-ray crystaIIography.
First a group of proteins called transcription
factors locate and bind to the TATA box.
Transcription factors - a collection of proteins
that help RNA poIymerase II recognize and
bind to the promoter.
Now, RNA polymerase is able to bind to the
promoter
RNA polymerase -the enzyme responsible for
transcribing the pre-mRNA strand.
Eukaryotes have three RNA polymerases.
Each of these polymerases transcribes a
different class of genes: (prokaryotes have
one RNA polymerase)
RNA !oIymerase I - Genes encoding ribosomaI
RNA
RNA poIymerase II - Genes encoding
messenger RNA
RNA !oIymerase III - Genes encoding transfer
RNA
Eukaryotes have several general transcription
factors (GTFs)- (prokaryotes only have sigma
factor)
RNA Pol does not contain a subunit similar to
the prokaryotic sigma factor, which can
recognize the promoter and unwind the DNA
double helix.
The RNA Pol is associated with six general
transcription factors, designated as TFA, TFB,
TFD, TFE, TFF and TFH, where "TF"
stands for "transcription factor" and "" for the
RNA Pol .
Aided by transcription factors, RNA
polymerase locates the start point and
begins to unwind the DNA helix.
RNA polymerase can then begin
transcribing the DNA sequence.
!art II- EIongation
RNA polymerase assembles the RNA
nucleotide that complement the template strand.
The enzyme reads the template strand in a 3' to
5' direction, while building the RNA strand in a 5'
to 3' direction.
RNA contains the base uracil (U) instead of
thymine (T) and is complementary to adenine
(A).
As RNA polymerase continues along the
transcription unit, the newly synthesized RNA
strand separates from the template.
3) !art III- termination
As the RNA transcript neras completion,
the enzyme encounters a terminator, or a
sequence of nucleotides that signals the
end of transcription.
The RNA polymerase transcribes the
terminator sequence, and then continues
for about 10-15 nucleotides before the pre
mRNA strand is released.
Before the pre-mRNA strand is released
from the nucleus, it must undergo
additional RNA processing.
This is done by modifying the ends of the
strand so that it will be protected from
being degraded in the cytoplasm.
5' cap
The 5' end of the strand is capped with
methylated guanosine
The 5' cap protects the mRNA from
degradation by hydrolytic enzymes as it
travels through the cytoplasm.
t also serves as a signal for the
attachment of the ribosome.
3' end
An enzyme called poly (A) polymerase
adds a tail of 100-00 adenine (A)
nucleotides to the 3' end of the mRNA.
The poly (A) tail protects the mRNA from
degradation and help in recognition by the
ribosome.
ntrons removed
The next step in the preparation of the
mRNA strand is the removal of the introns
(non coding segments).
Introns are sequences that are part of the
primary transcript but do not contribute
useful information for coding a protein.
Exons are the coding segments.
The end point of intron contain nucleotide
sequences that are recognized as splice
sites, where the strand is to be cut.
$pIiceosome locates the splice sites and
cuts out the intron.
Within the spliceosome are smaller RNA-
protein complexes called sn RN!s or
smalle nuclear ribonucleoproteins
ntrons are removed by multiple
spliceosome along the mRNA strand, and
the leftover exons join together.
Now the mRNA strand is ready to leave
the nucleus into the cytoplasm.
Translation of protein will begin in the
cytoplasm
Translation
Polypeptide is synthesized from the mRNA
strand using the genetic information
present in the mRNA.
During translation the nucleotide is
translated into amino acids.
A ribosome is an organelle comprised of
protein and rRNA (ribosomal RNA).
Ribosomes are the site of translation.
Part 1: nitiation
nitiation begins when the mRNA
transcript comes together with the
smaller of two ribosomaI subunits and a
tRNA moIecuIe.
The tRNA (transer RNA) carries the 1
st
amino acid (methionine)(UAC) of the
polypeptide.
The tRNA attaches to the start codon
(AUG) of the mRNA transcript.
The larger ribosomal subunit has 3 tRNA
binding sites (E, P and A)
The incoming tRNA binds to the A site.
The P site holds the tRNA that carries the
growing polypeptide strand.
The tRNA will exit the moleculevia E site
Part -Elongation
Divided into 3 steps.
Codon recognition
Bond formation
Translocation
Codon recognition: every 3 nucleotides
represent a codon which codes for a particular
amino acid.
The incoming tRNA contains an anticodon which
complements the codon in the A site.
Hydrogen bonds hold the nucleotides
together (codon and anticodon)
The amino acid will be joined by peptide
bond.
Then the tRNA together with the mRNA
template move down one site.
The used tRNA will be released via the E
site.
Part 3: termination
Translation is terminated when a stop
codon (UAA) is reached in the mRNA
transcript.
The release factor is a protein that
recognizes and binds to stop codon and
hydrolyze the chain, releasing the
polypeptide.