GENETIC REGULATION 2

MOLECULAR BIOLOGY
(WEEK 14)

LOOP FORMATION

lac OPERON ATTENUATION

Transcriptional termination regulation by secondary mRNA structure
Low tryptophan level
Attenuation
High tryptophan level

mRNA
mRNA

Ribosome is
stalled at trp
codon

Ribosome
No hairpin formation,
RNA polymerase
continues on attach
DNA
Attenuator
Leader
region is
completely
translated

Formation of this hairpin results

in the termination of transcription
as RNA polymerase stalls on
attached DNA

TRYPTOPHAN METABOLISM

ARABINOSE OPERON

DNA

RNA polymerase CAP binding site

araI

araO1

araO2

PC

PBAD

araC mRNA

araO2
araO1 araC

proteins

araO1 PBAD
CAP binding site
CAP-cAMP
RNA polymerase
PC

DNA
araO2

araO1 P
BAD
PC
CAP binding site
araBAD mRNA
Arabinose

TRANSLATIONAL
REGULATION:
AUTOGENOUS
CONTROL
KONTROL AUTOGENUS
(sintesis protein
ribosom)
DNA

S4 gene

Transcription

Ribosome

Polycistronic mRNA
Translation

S4 protein

When levels of ribosomal protein

S4 are high, they bind to mRNA
preventing further translation

TRANSLATIONAL REGULATION: RNA CONTROL

Synthesis of
extracellular-omp
protein

Lipopolysaccharide

OmpC, F,
NmpA, B, C

OmpA

Phospholipid

LamB

Lipoprotein

Peptidoglycan

micF gene
micF mRNA
complimenter
micF mRNA combine to OmpF
mRNA inhibit OmpF
translation no OmpF protein
incomplete OmP protein (only
OmpC protein)

MalE

OmpC gene
OmpC mRNA
OmpC protein
extracellular-Omp protein
OmpF protein
OmpF mRNA
OmpF gene

REGULATION BY CODON USAGE

one aa coded by several codons several type of tRNA for one aa
different tRNA population of each codon
high tRNA population of the use codon >> translation >> protein >>

REGULATION BY mRNA-SECONDARY STRUCTURE

AUG

dengan loop translasi <<

AUG

tanpa loop translasi >>

loop (mRNA-secondary structure) more difficult scanning by ribosome

looped RNA more stable stability >> energy << translation <<
most in eukaryote, in prokaryote, translation begins directly before the end of
transcription no mRNA-loop formation

REGULATION BY PHOSPHORYLATION
60S ribosomal subunit
mRNA
Methionyl initiator tRNA
Active
eIF-2

40S ribosomal
subunit

Protein
synthesis

Guanine nucleotide
releasing protein, eIF-2B

Inactive eIF-2

In the absence of
active eIF-2B, excess
eIF-2B
eIF-2 remains in its
Protein kinase
inactive, GDP-bound
phosphorylates eIF-2
form and protein
synthesis slows
dramatically
Phosphorylated eIF-2 sequesters all eIF-2 as an inactive complex

REGULATION OF GENE EXPRESSION IN EUKARYOTE

- GENERAL EXPRESSION FACTOR

- OTHER EUKARYOTIC REGULATION OF
GENE EXPRESSION

TRANSCRIPTIONAL REGULATION IN EUKARYOTE

GENERAL TRANSCRIPTION FACTORS
Start of transcription

TFIIF

TFIIE

RNA polymerase II

TFIIH
TFIID

Protein kinase (TFIIH) activity

TFIIB

Transcription begins
OTHER/LONG DISTANCE TRANSCRIPTION FACTOR
General transcription
Gene
factors
RNA
regulatory
polymerase II
proteins
Gene x
Regulatory Spacer DNA
sequences

TATA
Promotor

RNA transcript
The gene control region for gene x

Gene
regulatory
proteins

GENE REGULATION AT A DISTANCE: ENHANCER ELEMENT

DNA double helix

100 nucleotide pairs

500 nucleotide pairs
NtrC
Enhancer

Promotor

Looped
activation
intermediate
Gene on

GENE REGULATION AT A DISTANCE IN EUKARYOTE

Trans acting element/protein

Strong activating
assembly

Silent assembly of
regulatory proteins

Strongly
inhibiting
protein

Weakly
activating
protein
assembly

RNA polymerase and general

transcription factors

TATA

LONG DISTANCE TRANSCRIPTION-REGULATION MECHANISMS

Activator

Activation surface
Repressor

Competitive
DNA binding
Binding site
for activator

Binding site
for repressor

Masking the

activation surface

Binding site
for activator

TATA

TATA
Binding site
for repressor

Binding site
for activator

Direct interaction

with the general

transcription factor
TATA

GAL4-REGULATION
GAL4
protein

Acidic activation domain

GAL4 DNAlacZ gene
binding domain

DNA binding
site for GAL4

TATA

RNA

GAL4
TFIID

Gene on

lexA-GAL4
chimeric protein

TFIIB
TATA

Gene

DNA-binding
site for GAL4

TATA

Gene off

DNA-binding
site for lexA

TATA

Gene on
RNA