DNA Transcription & Translation

Dr. May Valzon

 How Cells Read the Genome: From DNA to
Protein?
The DNA in genomes does not direct protein synthesis
itself
uses RNA as an intermediary
When the cell needs a particular protein, they do:
a process called transcription
a process called translation
a principle so fundamental that it is termed the
central dogma of molecular biology
Flow of genetic information in cells is therefore from
DNA to RNA to protein
(central dogma of molecular biology)
 FROM DNA TO RNA
(DNA Transcription)
Portions of DNA sequence are transcribed into
RNA gen
Named transcription because still written in
essentially the same language as it is in DNA

Each transcribed segment of DNA is called a
transcription unit
Transcription produces RNA complementary
to one strand of DNA (template)
 First step is Opening and unwinding of a small
portion of the DNA
The RNA is elongated one nucleotide at a time
The RNAs nucleotide sequence have exactly
complementary to template
RNA strand doesnt hydrogen bond to the DNA
template strand
DNA transcription is a enzymatically catalyzed
reaction RNA polymerases
 RNA
RNA is a linear polymer made of
four different types of nucleotide
that linked by ribose sugar and
than linked together by
phosphodiester bonds
Have 4 Nitrogen bases: A, G, C, U
RNA is single-stranded that are
no more than a few thousand
nucleotides long
RNA can fold into specific
structures
Cells Produce Several Types of RNA
 RNA Polymerase
The enzym: RNA polymerases
RNA polymerases catalyze the formation of the
phosphodiester bonds that link the nucleotides together to
form a linear chain
The substrates are nucleoside triphosphates (ATP, CTP, UTP,
and GTP)
Work on 3-to-5 direction of DNA template produce 5-
3 elongated RNA
The RNA polymerase moves stepwise along the DNA
rate 20 nucleotides per second
RNA polymerases make about one mistake for every 104
nucleotides copied into RNA
RNA polymerases have a modest proofreading mechanism
RNA Pol Orientation
Transcription Process in Eucaryotic

Transcription Initiation

Transcription Elongation

RNA Processing
 Transcription Initiation
Signals Encoded in DNA Tell RNA Polymerase
Where to Start and Stop
In Bacteria are more simple: RNA polymerase
holoenzyme = RNA polymerase + sigma () factor
Promoter a special sequence of nucleotides
indicating the starting point for RNA synthesis
Terminator a special sequence of nucleotides
indicating the stoping point for RNA synthesis
a special sequence of nucleotides indicating the
starting point for RNA synthesis
Transcription initiation in Bacteria (procariotic)
 Transcription Initiation in Eucaryotes
In contrast to bacteria, which contain a single
type of RNA polymerase, eucaryotic nuclei
have three: RNA polymerase I, RNA
polymerase II, and RNA polymerase III
 RNA Polymerase II Requires General
Transcription Factors
general transcription factors function:
1. help to position eucaryotic RNA polymerase correctly at the
promoter
2. aid in pulling apart the two strands of DNA to allow transcription to
begin
3. release RNA polymerase from the promoter into the elongation
mode once transcription has begun
Initiation of Transcription
A. The promoter contains a DNA sequence called
the TATA box, which is located 25 nucleotides
away
B. from the site at which transcription is initiated.
C. Through its subunit TBP, TFIID recognizes and
binds the TATA box, which then enables the
adjacent binding of TFIIB
D. The rest of the general transcription factors, as
well as the RNA polymerase itself, assemble at
the promoter transcription initiation complex
E).
TFIIH contains a DNA helicase possible to
unwinding the DNA and hydrolyzing ATP.
TFIIH also phosphorylates RNA polymerase II,
changing its conformation so that the polymerase
is released from the general factors and can begin
the elongation phas of transcription.
the site of phosphorylation is a long C-terminal
polypeptide tail, also called the C-terminal
domain (CTD), that extends from the polymerase
molecule.
Gambaran Start Point Transkripsi pada Eukariotik
Polymerase II Also Requires Activator, Mediator,
and Chromatin-Modifying Proteins
 Transcription Elongation
RNA polymerase moves jerkily, pausing at some
sequences and rapidly transcribing through
others
Need a series of elongation factors proteins
that decrease the likelihood that RNA polymerase
will dissociate before it reaches the end of a gene
elongation factors also facilitate transcription
through nucleosomes without requiring
additional energy
 DNA supercoiling a conformation that DNA
adopts in response to superhelical tension
one largeDNA supercoil will form to compensate
for each 10 nucleotide pairs that are opened
(unwound)
this tension should facilitate the unwrapping of
DNAin nucleosomes, as the release of DNA from
the histone core helps to relax
positivesuperhelical tension
In eucaryotes, DNA topoisomerase enzymes
rapidly remove this superhelical tension
 RNA processing
Transcription Elongation in Eucaryotes Is
Tightly Coupled to RNA Processing
There are two RNA critical processing:
1. The covalent modification of the ends of the
RNA
Capping on the 5 end
Polyadenylation of the 3 end
2. Splicing removal of intron sequences that are
discarded from the middle of the RNA transcript
 RNA Capping Is the First Modification
of Eucaryotic Pre-mRNAs
As soon as RNA polymerase II has produced about 25
nucleotides of RNA, the 5 end of the new RNA
molecule is modified by addition of a cap that consists
of a modified guanine nucleotide
Three enzymes, acting in succession, perform the
capping reaction:
1. a phosphatase removes a phosphate from the 5 end
of the nascent RNA,
2. a guanyl transferase adds a GMP in a reverse linkage
(5 to 5 instead of 5 to 3), and
3. a methyl transferase adds a methyl group to the
guanosine
 RNA Splicing
RNA Splicing Removes Intron Sequences from
Newly Transcribed Pre-mRNAs
FROM RNA TO PROTEIN
(TRANSLATION)