The Central Dogma:

DNA replication, Transcription

and Translation
Blok VI FK UMP 2014

Yuwono, MD., PhD

Dept. Microbiology & Molecular Medicine
University of Sriwijaya
E-mail: yuwonodr@gmail.com
 Learning Objectives
Student:
1. Understand the mechanism of DNA
replication
2. Understand the mechanism of RNA
transcription
3. Understand the mechanism of Amino Acid
translation
4. Significance of central dogma in medicine
From cellular to molecular biology
 Cell Construction
Living cells: bioreactors with >2000
(mostly coupled) reactions

Proteins
Lipids
Carbohydrates
Nucleic acids
 Bio-molecules

• Nucleic acids (DNA, RNA): Library of life

• Proteins: Workhorse of life

• Fatty acids, carbohydrates, and other

supporting molecules
 Proteins Functions

• Structural
• Enzymes
• Information exchange (e.g., across cell walls)
• Transporting other molecules (e.g., oxygen to
cells)
• Activating-deactivating genes
• Etc.
(B) A polysaccharide molecule (red)—a polymer chain of sugar
monomers—binds to the catalytic site of lysozyme and is broken
apart, as a result of a covalent bond-breaking reaction catalyzed by
the amino acids lining the groove.
Life as an autocatalytic process.
Polynucleotides (nucleotide polymers) and proteins (amino acid
polymers) provide the sequence information and the catalytic
functions that serve—through a complex set of chemical reactions—
to bring about the synthesis of more polynucleotides and proteins of
the same types.
All cells store their genetic information in DNA

An egg cell.
The DNA of this single cell contains the genetic
information needed to specify construction of an entire
multi-cellular animal.
Hierarchy of DNA Structure
A gene is a segment of DNA that
encodes for (specifies) a protein
(via mRNA), a tRNA or an rRNA.

A genome refers to the total

complement of DNA in a cell.
Structure of Mammalian Genes
 The Central Dogma of Molecular Biology:
DNA------>RNA------>protein

The central dogma concerns the flow of biological

information: DNA is a self-replicating molecule containing
genetic information that can be transcribed into an RNA
message that can be translated into a polypeptide (protein).
 From DNA to Protein

Figure 6-21. Summary of the steps leading from gene to protein in eucaryotes and bacteria. The final level of a protein in the cell depends on the
efficiency of each step and on the rates of degradation of the RNA and protein molecules. (A) In eucaryotic cells the RNA molecule produced by
transcription alone (sometimes referred to as the primary transcript) would contain both coding (exon) and noncoding (intron) sequences. Before it can be
translated into protein, the two ends of the RNA are modified, the introns are removed by an enzymatically catalyzed RNA splicing reaction, and the
resulting mRNA is transported from the nucleus to the cytoplasm. Although these steps are depicted as occurring one at a time, in a sequence, in reality
they are coupled and different steps can occur simultaneously. For example, the RNA cap is added and splicing typically begins before transcription has
been completed. Because of this coupling, complete primary RNA transcripts do not typically exist in the cell. (B) In procaryotes the production of mRNA
molecules is much simpler. The 5 end of an mRNA molecule is produced by the initiation of transcription by RNA polymerase, and the 3 end is produced
by the termination of transcription. Since procaryotic cells lack a nucleus, transcription and translation take place in a common compartment. In fact,
translation of a bacterial mRNA often begins before its synthesis has been completed
The Central Dogma of molecular biology

Synthesis of three types of informational molecules: DNA, RNA

and Protein. Note that only one strand of the DNA is transcribed
into RNA.
DNA Replication is Semiconservative
DNA
replication
Is bidirectional
The replication fork -
where the action takes
place.

During replication, as the

DNA is unwound and the
two strands are separated,
the newly synthesized
strand that is oriented 5'--
>3' is called the leading
strand, and the new strand
that is oriented
3'-->5' is called the lagging
strand.
 Transcription Fundamentals

• Transcription is synthesis of RNA using a DNA template

• Three Key differences between DNA and RNA:
1. RNA contains the sugar ribose instead of deoxyribose
2. RNA contains the base uracil instead of thymine
3. Except in certain viruses RNA is not a doublestranded
molecule

Three major types of RNA - all products of transcription of DNA:

1. messenger RNA (mRNA)
2. transfer RNA (tRNA)
3. ribosomal (rRNA)
mRNA Reading Direction Corresponds to
Protein Chemical Directionality
mRNA
5’ 3’

NH2-terminus

COOH-terminus
Transcription is the process by which nucleotide
sequences in DNA are copied to a complementary copy of
messenger RNA
RNA polymerase is the
enzyme that copies DNA into a
complementary copy of RNA.

The enzyme uses DNA as a

template and since it catalyzes
the addition of ribonucleotides
in 5'-->3' direction, it reads its
template DNA in the 3'-->5'
direction.
Promoters
Promoters are specific sites
on DNA that RNA polymerase
first binds to to initiate the
transcription of a gene.
Sigma factors
Sigma factors are one
component of the
multicomponent RNA
polymerase enzyme that allow
the RNA polymerase to
recognize the initiation
(promoter) site.
Transcription Terminators

Transcription terminators are

sequences of nucleotide
bases at the end of the gene
that signal termination of
transcription.
Overview of Transcription
During translation, the genetic code in mRNA is read
and converted into protein by means of the protein
synthesizing machinery, which consists of ribosomes, tRNA,
amino acids, and a number of enzymes.
 The Genetic Code
allows for correspondence between triplets of bases in DNA
and the amino acid sequence of a polypeptide (protein)
written or expressed in terms of RNA triplets as compared to
DNA triplets because it is with messenger RNA that the
translation process occurs
 The Genetic Code
• Codons: Triplets of three bases in RNA that encode
an amino acid. There are 64 possible codons (4 bases
taken 3 at a time = 43)
• Stop and start codons:
Start = AUG (codes for methionine) - site where
translation begins
Stop = UAA, UAG and UGA - sites where translation
ends
• Degeneracy: Most amino acids have more than one
codon. For example, glycine is encoded by GGU GGC
GGA and GGG.
The structure of transfer RNA (tRNA)
 Steps in Protein Synthesis
1. initiation
• begins with an initiation
complex made up ribosomes,
initiation factors, mRNA and
formyl methionine tRNA. The
formyl methionine is delivered to
the initiation site on the mRNA,
codon AUG.

• There is a specific sequence of

bases on the mRNA, upstream
of the start codon, that allows
the ribosome to recognize and
bind onto the start site to initiate
the translation process.
2a. elongation
The mRNA is threaded through
the ribosome which contains
other sites where tRNAs
interact. The acceptor (A) site
is the site where the new
charged tRNA first attaches.
The peptide (P) site is the site
where a growing peptide is
held by a tRNA and where
peptide bond formation takes
place between the incoming
amino acid (at the A site) and
the amino acid at the P site.
2b. translocation
the ribosome advances by
three nucleotides exposing a
new codon at the A site and
pushing the now empty tRNA to
the exit (E) site where it is
released from the ribosome.
3. termination
occurs when a nonsense (stop)
codon is encountered. No tRNA
binds to a stop codon. Instead,
specific proteins called release
factors recognize this chain
terminating signal and cleave
the completed polypeptide from
the terminal tRNA.
Overview of Translation
Medical Significance
Cell Cycle
DNA Damages
SS breaking 55 000 per day & cell
Loss of purines12 000 per day & cell
Loss of pyrimidines 600 per day &
cell
Cytosine desamination 200 per day
& cell
DS breaking 9 per day & cell
Strand cross links 8 per day & cell
Pyrimidine dimer formation
rare
Mechanism of Gene Expression
 THREE MAIN TYPES OF
GENETIC DISEASE
• Mendelian : Determined by single major gene

• Multifactorial: Multiple genetic and non-genetic

factors involved

• Chromosomal: Multiple genes and visible

chromosomal abnormality
Queen Victoria’s Family and Hemophilia
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