GENE EXPRESSION

dr. Retno Sutomo, Ph.D, Sp.A

 Central dogma in molecular biology

DNA
transcription

RNA
translation

The flow of genetic information

Protein

"DNA makes RNA, RNA makes protein, and proteins make us“
(Francis Crick)
Crick’s
original
central dogma
The structure of DNA
The structure of DNA

Double-stranded, anti-parallel helix

 DNA structure

 DNA strands  complementary sequences/base

complementarity
 The sequence one DNA strand can be inferred if
that of its complementary strand is known
 Writing the DNA sequence  bases of one strand
only, in 5′ → 3′ direction

5’-GGTAGCTTAACCGAT-3’ GGTAGCTTAACCGAT
3’-CCATCGAATTGGCTA-5’
Bases in nucleic acids

Watson-Crick rule
 G -------- C
 A ---------T
Nucleotide nomenclature
Hydrogen bond between two bases
DNA REPLICATION

semi-conservative
 DNA replication

Okazaki fragments

only the leading strand is synthesized continuously

 semi-discontinuous
 RNA TRANSCRIPTION
 Transcription  synthesis of RNA
 DNA-dependent RNA polymerase
 In nucleus of eukaryotic cells and, to a limited extent, in
mitochondria and chloroplasts
 DNA is the hereditary material in all present-day cells
 Early in evolution, RNA served that function
 RNA can undergo self-replication but the sugar-phosphate
bonds of RNA are comparatively unstable chemically
 Many different classes of present-day viruses have a genome
that consists of RNA, not DNA
 RNA TRANSCRIPTION

 Retroviruses (ie. HIV)  a subclass of RNA viruses

 RNA replicates via a DNA intermediate, using a reverse
transcriptase (an RNA-dependent DNA polymerase)
 Eukaryotic cells, including mammalian cells, contain
nonviral chromosomal DNA sequences which
encode cellular reverse transcriptases
 Some nonviral RNA sequences act as templates for
cellular DNA synthesis  unidirectional flow of
genetic information is no longer strictly valid
 RNA TRANSCRIPTION
 Only a small proportion of all DNA in cells is transcribed
 Different cells transcribe different segments of the DNA
(transcription units)  discrete units, spaced irregularly
along the DNA sequence
 The great majority of the cellular DNA is never
transcribed in any cell
 Only a portion of the RNA made by transcription is
translated into polypeptide
 RNA molecule other than mRNA
 The primary transcript is subject to RNA processing events
 Only a central part of the mature mRNA is translated
 RNA TRANSCRIPTION
RNA is transcribed as a single strand
The RNA transcript sequence
 identical to the sense strand (except U replaces T)
 complementary to the template strand
RNA TRANSCRIPTION
Relationship among the three types of RNA
 RNA processing

 RNA splicing
 RNA capping
 RNA polyadenylation
RNA splicing
Consensus sequences at exon-intron junction
Mechanism of RNA splicing (GU-AG intron)
 Capping of mRNA

 Linking of 7-methylguanosine (m7G)

to the first 5′ nucleotide by a special
5′-5′ phosphodiester bond
 The 5′ end is blocked or capped

 To protect the transcript from 5′  3′  To facilitate RNA splicing

exonuclease attack  To facilitate attachment of the 40S
 To facilitate transport from the subunit of the cytoplasmic
nucleus to the cytoplasm ribosomes to the mRNA
 Poyadenylation

Poly (A) tail

 To facilitate transport of the mRNA molecules to the cytoplasm

 To stabilize at least some of the mRNA molecules in the
cytoplasm
 To facilitate translation by permitting enhanced recognition of
the mRNA by the ribosomal machinery
 TRANSLATION

 mRNA decoded on ribosomes to specify the

synthesis of polypeptides
 Post-transcriptional processing  mRNA migrates
to the cytoplasm  engages with ribosomes and
other components to direct the synthesis of specific
polypeptides
 The mitochondria also have ribosomes and a
limited capacity for protein synthesis
 TRANSLATION

 Only the central segment of mRNA is translated

to specify the synthesis of a polypeptide
 5′ and 3′ untranslated regions (5′ UTR; 3′ UTR),
 assist in binding and stabilizing the mRNA on
the ribosomes where translation of the central
segment occurs
• A group of 3 nucleotides in RNA constitute coding unit (codon)
• Each codon specifies a particular amino acid
• 4 bases  64 (43) possible codon combinations
• In fact only 20 AA specified  each specified by ≥1 codon

genetic code is degenerate

• Some AA have only one codon, others have up to 6 codons

• Codon AUG codes for methionine  start codon
• Codon UAA, UAG and UGA  termination (stop) codons.
Translation of genetic codes
• 64 codons in mRNA  30 cytoplasmic tRNA
22 mitochondrial tRNA
• How can all 64 codons interpreted?
– Normal base-pairing rules are relaxed in codon-anticodon
recognition
– Pairing of codon and anticodon follows the normal A-U and G-
C rules for the first two base positions in a codon
– Exceptional ‘wobbles' occur at the third position

wobble hypothesis
Wobble hypothesis
TRANSLATION

Translation of messenger RNA into protein product

 codon-anticodon recognition

 mRNA sequence is interpreted from start codon until a stop codon in 5′ → 3′

direction
 Each codon in mRNA is recognized by the complementary anticodon
sequence in tRNA
 mRNA sequence is interpreted from a translational start point (marked by the
sequence AUG) and continues in the 5′ → 3′ direction until a stop codon is
reached in that reading frame
 Each codon in the mRNA is recognized by the complementary anticodon
sequence of a tRNA molecule
Post-translational modification
 Addition of specific chemical groups to
specific amino acids
 Protein modification by addition of carbohydrate
groups
 Protein modification by addition of lipid groups
 Post-translational cleavage
 Human insulin

Multiple post-translational cleavages of polypeptides precursor

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