S-BIOL327A – INTRODUCTION TO GENETICS Lecture

1ST SEM (2024-2025) – Period 1st Sem 2024-2025

Finals: Module 7 Central Dogma of Molecular Biology

Schoolbook: DNA Replication

DNA replicates in a semi-conservative

manner. The two strands of the parent
DNA serve as the template or pattern to
which a complementary strand is
synthesized.

The accuracy of replication is contributed

by specific base pairing and proofreading
activities of the enzymes involved.
The central dogma of molecular biology
consists of three processes namely
Replication, Transcription, and
Translation.

DNA replicates in semi-conservative mode

that is, a newly synthesized molecule
contains an old strand from the parent

DNA and a newly synthesized DNA strand.

The two strands of the parent DNA serve as
the template or pattern to which a
complementary strand is synthesized. The
accuracy of replication is guaranteed by
specific base pairing.

In Transcription process, RNA is being

synthesized from the DNA template. It
comprises three general steps: The process begins with the separation of
the double strand (ds) DNA into single
1) Initiation
strands (ss) that would serve as the
2) Elongation template.

3) Termination The enzyme needed is the helix unwinding

protein (HUP) which uses ATP to unwind the
Three types of RNA dsDNA by breaking the hydrogen bonds in
between nucleotide pairs. To keep the
1. messenger RNA single strands from reannealing, several
2. transfer RNA helix destabilizing proteins (HDP), also
3. ribosomal RNA known as single strand binding protein
They have their own specific function. (SSBP) bind to the strands of the DNA.

In Translation process, proteins are A third enzyme is the helix relaxing protein
synthesized. A polypeptide chain from the (HRP) or the DNA gyrase relaxes twisting
activation of amino acids, initiation and tension (supercoils) created during
termination of translation to undergo unwinding. Once the templates are open,
further processing becomes a functional short segments of RNA nucleotides are
protein. synthesized by RNA polymerase to serve as
primers for the elongation of the (exonuclease activity) in 3’ to 5’
complementary strand. direction. It then replaces the excised
primers with DNA nucleotides but leaves a
RNA primers provide the 3’OH needed by
break between the DNA segments.
the DNA III to catalyze the phosphodiester
bond when nucleotides are added during The gaps or breaks within the newly
elongation. The leading strand needs only synthesized strands are sealed or joined by
one primer because it opens its an enzyme called ligase. Eventually, the
complementary strand in a 5’ to 3’ whole DNA has been replicated faithfully.
direction.

The other strand is designated as the

lagging strand opens its complementary
strand in a 3’ to 5’ direction. Several primers
are needed when the complementary
strand elongates in lagging strand.

Gene Action and Expression

Transcription and translation between

prokaryotes and eukaryotes differ mainly
due to their cellular structures. In
eukaryotes, transcription occurs within the
nucleus and the RNA products are then
transported to the cytoplasm for protein
synthesis.
Once the primers are synthesized, the DNA
polymerase III is capable of adding Prokaryotes, in absence of nucleus,
nucleotides by covalently bonding it from synthesize their RNA and proteins in the
the primer. cytoplasm.

Nucleotides are added as nucleotide Transcription

triphosphates such as ATP, GTP, TTP, CTP
➢ The RNA polymerase attaches to a
that also serve as energy source during
specific sequence in the DNA called
bonding. The complementary strand in
the promoter sequence or the
both templates elongates from 5’ to 3’
initiation point with the help of the
direction. The leading strand elongates
polypeptide called the sigma factor
continuously. This is because the added
(ð).
nucleotide in the leading strand provides
➢ Sigma factor recognizes the
the available 3’OH needed for covalent
promoter sequence so that the RNA
bonding as the strand elongates from 5’ to
polymerase can selectively bind to
3’. Its direction is towards the unwinding of
it. Once bound, ð dissociates and is
DNA. The lagging strand on the other
free for another round of initiation.
hand, elongates in a discontinuous
➢ The RNA polymerase moves along
manner.
one of the strands of the DNA called
The lagging strand is oriented away from the anticoding strand or antisense
the unwinding region strand eventually strand, it complements the DNA
contains short DNA segments (~1000-2000 nucleotides with RNA nucleotides.
nucleotides), known as Okazaki fragments, ➢ When the segment of DNA is
separated by RNA primers. Another completely transcribed, the DNA
enzyme, DNA polymerase I proofreads the strands reanneal because the DNA-
DNA and removes the primers by digestion DNA interaction is stronger than
 DNA-RNA interaction. A single ➢ The fMET-tRNA complements the
stranded RNA is thus produced. This AUG codon at the 5’end of the
continues until the enzyme reaches mRNA. This site in the ribosome is
the termination signal. termed as the P-site (peptidyl
➢ Another protein factor called Rho binding site). The other site which is
factor binds and dissociates from the responsible for the entry of other aa-
DNA template to prevent further tRNA is the Asite (amino-acyl site).
transcription. Another type of ➢ Once the fMET is set at the P-site, a
termination is by loop formation on new aa-tRNA complex enters the A-
the GC rich sequence of the site complementary to the
synthesized RNA strand which pulls it succeeding codon. An enzyme,
away from the DNA template. peptidyl transferase, present in the
➢ There are three different types of 50s subunit catalyzes a peptide
RNAs namely: messenger RNA bond between the fMET and the
(mRNA), transfer RNA (tRNA) and amino acid carried by the tRNA in
ribosomal RNA (rRNA) which are the A-site.
essential to the next process called ➢ The tRNA in the P-site is then released
translation. to the cytoplasm for
reactivation. This is followed by the
movement of the mRNA along the
ribosome that displaces the aa-tRNA
from the A-site to the P-site leaving
the A-site free for other incoming aa-
tRNA complex.
➢ Elongation is from 5’ to 3’ relative to
the mRNA. Termination of
Translation translations occurs when the
➢ The amino acids are attached to ribosome encounters one of the
their respective tRNAs. This requires termination codons (UAG, UGA,
energy from ATP, and it is mediated UAA). These codons do not code for
by amino-acyl synthetase any amino acid, but instead code
enzyme. The initiation codon AUG is for release factor proteins (RF1 or
always oriented on the 5’ end of the RF2). These factors dissociate the
mRNA. polypeptide and the tRNA from the
➢ Initial steps involve the attachment ribosome and eventually dissociates
of the first aa-tRNA complex with a the 70s ribosome into its subunits.
formylated methionine (fMET) to the ➢ The polypeptide chain can be a
30s subunit through the action of protein or undergoes further
initiation factor 1 (IF1). processing to become a functional
➢ At the same time, a protein IF3, binds protein.
the mRNA to 30s subunit and orients
the fMET to the AUG codon. IF2 then
completes the 70s ribosome by
binding the 50s to the 30s
subunit. GTP serves as the energy
source during these reactions. The
formyl group in fMET prevents the
elongation of the polypeptide at the
amino-end (N-terminal).
The Genetic Code

The genetic code consists of the sequence

of nitrogen bases—A, C, G, U—in an mRNA
chain. The four bases make up the “letters”
of the genetic code. The letters are
combined in groups of three to form code
“words,” called codons.

Each codon stands for (encodes) one

amino acid, unless its codes for a start or
stop signal. There are 20 common amino
acids in proteins. There are 64 possible
codons, more than enough to code for the
20 amino acids. The genetic code has a
few important characteristics.

• The genetic code is universal. All

known living organisms use the same
genetic code. This shows that all
organisms share a common
evolutionary history.

• The genetic code is unambiguous.

Each codon codes for just one
amino acid (or start or stop). What
might happen if codons encoded
more than one amino acid

• The genetic code is redundant. Most

amino acids are encoded by more
than one codon.