Features Prokaryotic Replication Eukaryotic Replication

Location Cytoplasm Nucleus

Timing Anytime before cell division S-Phase of the cell cycle

Rate Faster (2000 bp/s) Slower (100 bp/s)

DNA gyrase, DNA Helicase, Topoisomerase, DNA

Enzymes
Polymerase III Polymerases α, β, ε, γ

Replicons One Multiple

Ori Sites One Multiple

Terminus One Multiple

Okazaki Longer (1000-2000

Shorter (100-200 Nucleotides)
Fragments Nucleotides)

Accuracy More accurate Less accurate

DNA Replication

In the process of DNA replication, the DNA makes multiple copies of itself. It is a biological
polymerisation, which proceeds in the sequence of initiation, elongation, and termination. It is an
enzyme-catalysed reaction. DNA Polymerase is the main enzyme in the replication process.
DNA Replication Process

DNA Replication Steps

Following are the important steps involved in DNA replication:

Initiation

DNA replication demands a high degree of accuracy because even a minute mistake would result
in mutations. Thus, replication cannot initiate randomly at any point in DNA.

For the replication to begin there is a particular region called the origin of replication. This is the
point where the replication originates. Replication begins with the spotting of this origin
followed by the unwinding of the two DNA strands.

Unzipping of DNA strands in their entire length is not feasible due to high energy input. Hence,
first, a replication fork is created catalysed by the helicase enzyme, which unzips the DNA
strand.

Elongation

As the strands are separated, the polymerase enzymes start synthesising the complementary
sequence in each of the strands. The parental strands will act as a template for newly
synthesising daughter strands.

It is to be noted that elongation is unidirectional i.e. DNA is always polymerised only in the 5′ to
3′ direction. Therefore, in one strand (the template 3‘→5‘) it is continuous, hence called
continuous replication while on the other strand (the template 5‘→3‘) it is discontinuous
replication. They occur as fragments called Okazaki fragments. The enzyme called DNA ligase
joins them later.
DNA Replication Fork

Termination

Termination of replication occurs in different ways in different organisms. In E.coli like

organisms, chromosomes are circular. And this happens when the two replication forks between
the two terminals meet each other.

Role of Enzymes in DNA Replication

DNA replication is a highly enzyme-dependent process. There are many enzymes involved in
DNA replication, which includes the enzymes, DNA-dependent DNA polymerase, helicase,
ligase, etc. Among them, DNA-dependent DNA polymerase is the main enzyme.

DNA-dependent DNA polymerase

It helps in the polymerisation, catalyses and regularises the whole process of DNA replication
with the support of other enzymes. Deoxyribonucleoside triphosphates are the substrate as well
as the energy provider for the replication process. DNA polymerase is of three types:

DNA Polymerase I

It is a DNA repair enzyme. It is involved in three activities:

 5′-3′ polymerase activity

 5′-3′ exonuclease activity

 3′-5′ exonuclease activity

DNA Polymerase II

It is responsible for primer extension and proofreading.

DNA Polymerase III

It is responsible for in vivo DNA replication.

Helicase

Helicase is the enzyme, which unzips the DNA strands by breaking the hydrogen bonds between
them. Thus, it helps in the formation of the replication fork.

Ligase

Ligase is the enzyme which joins together the Okazaki fragments of the discontinuous DNA
strands.

Primase

This enzyme helps in the synthesis of RNA primer complementary to the DNA template strand.

Endonucleases

These produce a single-stranded or a double-stranded cut in a DNA molecule.

Single-stranded Binding Proteins

It binds to single-stranded DNA and protects it from forming secondary structures.

DNA Replication Process in Prokaryotes

The DNA replication in prokaryotes takes place in the following place:

1. The two strands of DNA unwind at the origin of replication.

2. Helicase opens the DNA and replication forks are formed.

3. The DNA is coated by the single-strand binding proteins around the replication fork to
prevent rewinding of DNA.

4. Topoisomerase prevents the supercoiling of DNA.

5. RNA primers are synthesised by primase. These primers are complementary to the DNA
strand.

6. DNA polymerase III starts adding nucleotides at the end of the primers.

7. The leading and lagging strands continue to elongate.

 8. The primers are removed and the gaps are filled with DNA Polymerase I and sealed by
ligase.

DNA Replication in Eukaryotes

The DNA replication in eukaryotes is similar to the DNA replication in prokaryotes. However,
the initiation process is more complex in eukaryotes than prokaryotes. In eukaryotes, there are
multiple origins of replication present. A pre-replication complex is made with other initiator
proteins. The process is entirely the same but the enzymes used are different. E.g. in eukaryotes,
the polymerisation process is carried out by the enzyme Pol δ, whereas in prokaryotes it is done
by DNA Pol III.

TRANSCRIPTION:

Eukaryotic Transcription Prokaryotic Transcription

Eukaryotic Transcription takes place inside of the Nucleus. Prokaryotic Transcription takes place in the
Cytoplasm.

In eukaryotic Transcription, combinational translation and In Prokaryotic Transcription, combinational

transcription are not possible. translation and transcription are possible.

In Eukaryotic Transcription, proteins known as In Prokaryotic Transcription, no such

Transcriptional factors are needed. proteins are needed.

The process of transcription carries three enzymes. These The process of transcription carries just one
enzymes are- RNA Polymerases I, RNA Polymerases II and enzyme. This enzyme synthesises all other
RNA Polymerases III. types of RNA.

This process includes five polypeptides. This process includes ten to fifteen
polypeptides.

Conclusion

Transcription is the first step of Gene Expression. Transcription is the process through which the
segments of DNA are copied into the RNA of a body. DNAs are used as a template for making
RNA molecules. The prokaryotes and the eukaryotes hold similar procedures of transcription.
The transcription process takes place in three stages. These stages are initiation, elongation, and
termination. The process of transcription in eukaryotes initiates in the nucleus. The mRNA
travels out of the nucleus and moves to the cytoplasm, where the process of translation takes
place. The process of transcription in prokaryotes initiates in the cytoplasm. RNA processing is
not required in prokaryotes. This is because the structural gene in prokaryotes is polycistronic in
nature. This process of transcription is also known as bacterial transcription.

TRANSALTION:

Prokaryotic Translation Eukaryotic Translation

Definition

The translation & transcription process is The translation and transcription process is
synchronous discontinuous

mRNA

Cytoplasm Nucleus

Cap initiation

Cap-independent Cap-dependent and Cap-independent

Performed by

70S ribosomes 80S ribosomes

Start site

Shine Dalgona One site towards 5 site of mrna

No shine dalgona seq

Ribosomes

30S & 50S = 70S 40S & 60S = 80S

First amino acid

fMET Methionine

Occurrence

No definite phase G1 and G2 phases of the cell cycle

mRNA first attached with rRNA of smaller subunit

16s 40s

Initiation factor

IF1, IF2, IF3 eIF