Expression of Genetic Information

First insight into gene function

Archibald Garrod (1908) - certain inherited

diseases were caused by absence of specific
enzymes which is coded by genes (as with other
cases, “Yaaawn... Zzzz..” was the response)

Beadle and Tatum (1940) resurrected the idea in

their studies of the bread mold, Neurospora
Expression of Genetic Information
Induce mutation
(irradiation) that
resulted in lack of a
particular enzyme
(infered by growth on
minimal medium)

“ONE GENE - ONE

ENZYME” (not true anymore..)
Expression of Genetic Information
Overview:

Transcription to mRNA

mRNA processing (euk)

Transport to cytoplasm

Translation (ribosomes,
tRNA)
Expression of Genetic Information

Transcription only
utilises info on one
strand
 Expression of Genetic Information
Transcription occurs locally aka transcription bubble
(unwind at one end and rewind at the other)
Expression of Genetic Information
RNA polymerase
catalyses the reaction

RTPn + NPPP  RMPn+1 + PPi

Expression of Genetic Information
Transcription stages

Initiation

Elongation

Termination
Expression of Genetic Information
Transcription stages
Transcription: Synthesis of RNA

• Enzymatic synthesis of RNA:

in vivo and in vitro

• DNA-dependent RNA polymerase

• RNA-dependent RNA polymerase

Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
Transcription: Synthesis of RNA

• Enzymatic synthesis of RNA:

in vivo and in vitro

• DNA-dependent RNA polymerase

• RNA-dependent RNA polymerase

Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
DNA-dependent RNA polymerase
• Prokaryotic system (E. coli)
• Complex enzyme: holoenzyme 
sigma (σ) factor and core enzyme
(α2ββ’ ─ three different polypeptide
chains)
• Sigma (σ) factor: recognizes the
transcription initiation signal
• Core enzyme: catalyzes the formation
of the internucleotide 3’-5’
phosphodiester bonds
Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
 Synthesis of RNA

n ATP ssDNA
n CTP template
n GTP
n UTP + 3’-AAGCTTTAGGGCTAACCG-5’
all four
Mg++
ribonucleoside
triphosphates
DNA-dependent RNA polymerase
5’-3’ extension, sequence
complementary to that of
the DNA template pyrophosphates, PPi

5’-UUCGAAAUCCCGAUU-3’ inorganic
3’-AAGCTTTAGGGCTAACCG-5’ phosphates, Pi
DNA-RNA hybrid duplex
Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
 Synthesis of RNA in vivo

DNA
5’-TTCGAAATCCCGATTGGC-3’ Non-template strand
3’-AAGCTTTAGGGCTAACCG-5’ template strand

transcription

5’-TTCGAAATCCCGATTGGC-3’
5’-UUCGAAAUCCCGAUUGGC-3’ Sense RNA strand
3’-AAGCTTTAGGGCTAACCG-5’ antisense strand
template DNA strand

Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
 Core enzyme

Sigma factor

5’ 3’
3’ 5’

upstrea
downstream
m

Consensus TTGACA TATAAT +1

-35 sequence -10 sequence
Pribnow sequence, box

promoter sequence
Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
 Synthesis of RNA from RNA template

n ATP ssRNA
n CTP template
n GTP
n UTP + 3’-AAGCUUUAGGGCUAACCG-5’
all four
Mg++
ribonucleoside
triphosphates
RNA-dependent RNA polymerase
RNA replicase, RNA synthetase

pyrophosphates, PPi

5’-UUCGAAAUCCCGAUU-3’ inorganic
3’-AAGCTTTAGGGCTAACCG-5’ phosphates, Pi
RNA-RNA duplex
Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
Expression of Genetic Information
Where does transcription start (beginning of a gene)?
Promoter - binding of polymerase and sigma factor

Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
Expression of Genetic Information
Where does transcription start (beginning of a gene)?
Promoter - binding of polymerase and sigma factor

 Elongation

Courtesy: Prof. Dr. Koh Chong Lek & Prof. Dr. Sam Choon Kook
Expression of Genetic Information
Termination
Type I - intrinsic factors
that induces hairpin
formation, that stalls the
polymerase
Intrinsic factors
inverted repeats
A/U rich region
Expression of Genetic Information
Termination
Type II - involvement of
rho factors