PROJECT REPORT

ON

DNA TRANSCRIPTION &

TRANSLATION

SUBMITTED BY: SUBMITTED TO:

PALAK CHAUDHARY NAVEEN DHIMAN
CLASS- XII DEPT. OF BIOLOGY
ROLL NO.-

RANA INTERNATIONAL SCHOOL,

DEGANA JN.
 CERTIFICATE

This is to certify that palak chaudhary, student of class XII

of this school has completed her project file under my

supervision. She has taken proper care and shown utmost

sincerely in completion of this project. I further certify that

this project is up to my expectations and as per guidelines

issued by Central Board of Secondary Education.

EXAMINER’S SIGNATURE

PRINCIPAL’S SIGNATURE

TEACHER’S SIGNATURE
 ACKNOWLEDGMENT

I would like to express my special thanks of gratitude to my

teacher Mr. Naveen Dhiman who gave me the golden

opportunity to do this wonderful project on the topic DNA

TRANSCRIPTION & TRANSLATION, which also helped

me in doing a lot of Research and I came to know about so

many new things. I am really thankful to them.

Secondly, I would also like to thank my parents and friends who

helped me a lot in finalizing this project within the limited time

frame.

PALAK CHAUDHARY
 INDEX

 INTRODUCTION

 TRANSCRIPTION

 TRANSCRIPTION UNIT

 PROCEDURE (TRANSCRIPTION)

O ACTIVATION OF tRNA

 INITIATION

 ELONGATION

 TERMINATION

 TRANSLATION

 MECHANISM OF PROTEIN SYNTHESIS

 INITIATION

 ELONGATION

 TERMINATION
 INTRODUCTION

Central dogma of life:

DNA structure:

 One monomer unit=deoxyribonucleic acid.

 Composed of a base, a sugar (deoxyribose), and a phosphate.
 Directionality along the backbone 5’ (phosphate) to 3’ (OH) Double-
strand pairing.
 Complementary base-matching: A-T, C-G.
 Base-matching achieved by H-bonding and geometry (long vs short
nucleotides).
 Antiparallel (one strand 5’3’, the other 3’5’) Helical shape.
 10.4 nucleotides per turn.
 Diameter = 2 nm.
 Both major and minor grooves.
 A, G are long (double ring purines).
 C, T are short (single ring pyrimidines).
 Need one long and one short nucleotide per pair.
 C-G have three hydrogen bonds (slightly stronger matching).
 A-T have two hydrogen bonds (slightly weaker matching).
 Base stacking of aromatic rings allows sharing of pi electrons and
adds stability to interior structure of DNA some hydrophobic driving
force as well.
 Pair structure allows template for semi-conservative copying.
Information in DNA sequence is the genome.
 Genes are stretches of information in the sequence that encode for
particular function (usually a particular protein, but sometimes also an
RNA sequence).
 About 20,000 genes in humans.
 Typically, 1000s of nucleotides long.
 Genes can be expressed (use to make proteins) or repressed (not
used).
 Regions of DNA are divided into coding and non-coding segments.
 Over 50% of human DNA is non-coding.
 Genes can be spliced together.
 Genes are organized in the large-scale structure of the DNA in the
nucleus.
 In bacteria, genome usually circular.
 The genome in eukaryotes is organized into chromosomes.
 Each chromosome a separate DNA molecule.
 TRANSCRIPTION: Synthesis of RNA

Transcription is a process of making a copy of genetic information stored in

a DNA strand into a complementary strand of RNA with the aid of RNA
polymerase.
 TRANSCRIPTION UNIT

Transcription unit in DNA has three regions:

[1] PROMOTER

[2] STRUCTURAL GENE

[3] TERMINATOR

 The promoter site is located towards 5’-end of the structural gene.

 The two strands of the DNA in the structural gene have oppositely

polarity. One of the two strands of the DNA in the structural gene that

has the polarity 3’-5’ acts as a template and referred to as template

strand. The other strand that has the polarity 5’-3’ acts as a coding

strand.

 5’ and 3’ end of structural gene referred with respect to the polarity of

coding strand. Thus, the promoter is located towards the 5’ end of

coding strand and the terminator is located towards the 3’ end of

coding strand.

 In fact, the enzyme RNA polymerase binds towards he promotor site

and starts transcription moving towards terminator.
 PROCEDURE
[1] INITIATION:
To begin transcribing a gene, RNA polymerase binds to the DNA of the
gene at a region called promoter. Basically, the promoter tells the
polymerase where to sit down on the DNA and begin transcribing.

Each gene has its own promoter. A promoter contains DNA sequences that
let RNA polymerase or its helper proteins attach to the DNA. Once the
transcription bubble has formed the polymerase can start transcribing.

Promoters in human
In eukaryotes like humans, the main RNA polymerase in our cells does not
attach directly to promoters like bacterial RNA polymerase. Instead, helper
proteins called basal transcription factors bind to the promoter first, helping
the RNA polymerase in our cells get a foothold on the DNA.
Many eukaryotes promoters have a sequence called a TATA box. The
TATA box plays a role much like that of the -10 elements in bacteria. It’s
recognized by one of the general transcription factors, allowing others
transcription factors and eventually RNA polymerase to bind. It also
contains lots of As and Ts, which make it easy to pull the strands of DNA
apart.

[2] ELONGATION:
Once RNA polymerase is in position at the promoter, the next step of
transcription –elongation- can begin. Basically, elongation is the stage when
the RNA strand gets longer, thanks to the addition of new nucleotides.

During elongation, RNA polymerase “walks” along one strand of DNA,

known as the template strand, in the 3’ to 5’ direction. For each nucleotide in
the template, RNA polymerase adds a matching RNA nucleotide to the 3’
end of the RNA strand.

The RNA transcript is nearly identical to the non-template, or coding, strand

of DNA. However, RNA strands have the base uracil(u) in place of
thymine(T), as well as a slightly different sugar in the nucleotides. So, as we
can see in the above diagram, each T of the coding strand is replaced with a
U in the RNA transcript.
[3] TERMINATION:

RNA polymerase will keep transcribing until it gets signals to stop. The
process of ending transcription is called termination, and it happens once the
polymerase transcribes a sequence of DNA known as a terminator.

Termination in bacteria:
There are two major termination strategies found in bacteria:
1. Rho-dependent
2. Rho-independent

In Rho-dependent termination, the RNA contains a binding site for a

protein called Rho factor. Rho factor binds to this sequence and start
“climbing” up the transcript towards RNA polymerase.
When it catches up with the polymerase at the transcription bubble, Rho
pulls the RNA transcript and the template DNA strand apart, realsing the
RNA molecules and ending transcription. Another sequence found late in the
DNA called the transcription stop point, causes RNA polymerase to pause
and thus helps Rho catch up.

Rho-independent termination:
Depends on specific sequence in the DNA template strand. As the RNA
polymerase approaches the end of the gene being transcribed, it hits a region
rich in C and G nucleotides. The RNA transcribed from this region folds
back on itself, and the complementary
C and G nucleotides bind together. The result is a stable hairpin that causes
the polymerase to stall.
In a terminator, the hairpin is followed by a stretch of U nucleotides in the
RNA, which match up with A nucleotides in the template DNA. The
complementary U-A region of the RNA transcript forms only a weak
interaction with the template DNA.
This, coupled with the stalls polymerase, produces enough instability for the
enzyme to fall off and liberate the new RNA transcript.
 TRANSLATION

Translation is the mechanism by which the triplet base sequence of mRNA

guides the linking of a specific sequence of amino acid to form a polypeptide
on ribosomes. Ribosomes has one binding sites for mRNA and three binding
sites for tRNA:
1. Psite (peptidyl-tRNA site)
2. A site (acceptor site) which holds the tRNA carrying next amino acid to
be added to the chain.
3. Esite (exit site) where discharged tRNA leave the ribosomes.
The amino acids are joined by a bond which is known as peptide bond.
Formation of peptide bond requires energy. Therefore, in the first phase
itself amino acids are activated in the presence of ATP and linked to their
cognate tRNA – a process commonly called as charging of tRNA. If two
such charged tRNAs are brought close enough, the formation of peptide
bond between them would be favored energetically.
 MECHANISM OF PROTEIN SYNTHESIS:

Translation involves three steps:

 Initiation
 Elongation
 Termination

Translation begins with the binding of the small ribosomal subunit to a

specific sequence on the mRNA chain. The small subunit binds via

complementary base pairing between one of its internal subunits and the

ribosome binding site, a sequence of about ten nucleotides on the mRNA

located anywhere from 5 and 11 nucleotides from the initiating codon, AUG.
 1. Initiation

Once the small subunit has bound, a special tRNA molecule, called N-
formyl methionine, or fMet, recognizes and binds to the initiator codon.
Next, the large subunit binds, forming what is known as the initiation
complex. With the formation of the initiation complex, the fMet-tRNA
occupies the P site of the ribosome and the A site is left empty. This entire
initiation process is facilitated by extra proteins, called initiation factors that
help with the binding of ribosomal subunits and tRNA to the mRNA chain.
 2. Elongation:

With the formation of the complex containing fMet-tRNA in the peptidyl

site, an aminoacyl tRNA with the complementary anticodon sequence can
bind to the mRNA passing through the acceptor site. This binding is aided
by elongation factors that are dependent upon the energy from the hydrolysis
of GTP. Elongation factors go through a cycle to regenerate GTP after its
hydrolysis.

Now, with tRNA bearing a chain of amino acids in the p site and tRNA
containing a single amino acid in the A site, the addition of a link to the
chain can be made. This addition occurs through the formation of a peptide
bond, the nitrogen-carbon bond that forms between amino acid subunits to
form a polypeptide chain. This bond is catalyzed by the enzyme peptidyl
transferase.

Peptide Formation
The peptide bond occurs between the carboxyl group on the lowest link in
the peptide chain located at the p site and the amine group on the amino acid
in the A group. As a
result, the peptid e chain shifts over to the A site, with the original amino
acid on the A site as the lowest link in the chain. The tRNA in the A site
becomes peptidyl RNA, and shifts over to the P site. Meanwhile, the
ribosome engages in a process called translocation: spurred by elongation
factors, the ribosome moves three nucleotides in the 3' prime direction along
the mRNA. In other words, the ribosome moves so that a new mRNA codon
is accessible in the A site.
Translocation:
With the A site open again, the next appropriate aminoacyl tRNA can bind
there and the same reaction takes place, yielding a three-amino acid peptide
chain. This process repeats, creating a polypeptide chain in the P site of the
ribosome. A single ribosome can translate 60 nucleotides per second. This
speed can be vastly augmented when ribosomes link up to form
polyribosomes.
Termination:

Translation ends when one of three stop codons, UAA, UAG, or UGA,
enters the A site of the ribosome. There are no aminoacyl tRNA molecules
that recognize these sequences. Instead, release factors bind to the P site,
catalyzing the release of the completed polypeptide chain and separating the
ribosome into its original small and large subunits.
 BIBLIOGRAPHY

 NCERT BIOLOGY

 WIKIPEDIA

 PRADEEP’S BIOLOGY

 GOOGLE