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Chapter 18

The Genetics of Viruses


and Bacteria

PowerPoint Lectures for


Biology, Seventh Edition
Neil Campbell and Jane Reece

Lectures by Chris Romero


Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• Overview: Microbial Model Systems
• Viruses called bacteriophages
– Can infect and set in motion a genetic takeover
of bacteria, such as Escherichia coli

0.5 m
Figure 18.1
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• E. coli and its viruses
– Are called model systems because of their
frequent use by researchers in studies that
reveal broad biological principles

• Beyond their value as model systems


– Viruses and bacteria have unique genetic
mechanisms that are interesting in their own
right

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• Recall that bacteria are prokaryotes
– With cells much smaller and more simply
organized than those of eukaryotes

• Viruses
– Are smaller and simpler still
Virus

Bacterium
Animal
cell

Animal cell nucleus


0.25 m
Figure 18.2
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• Concept 18.1: A virus has a genome but can
reproduce only within a host cell
• Scientists were able to detect viruses indirectly
– Long before they were actually able to see
them

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The Discovery of Viruses: Scientific Inquiry
• Tobacco mosaic disease
– Stunts the growth of tobacco plants and gives
their leaves a mosaic coloration

Figure 18.3

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• In the late 1800s
– Researchers hypothesized that a particle
smaller than bacteria caused tobacco mosaic
disease

• In 1935, Wendell Stanley


– Confirmed this hypothesis when he crystallized
the infectious particle, now known as tobacco
mosaic virus (TMV)

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Structure of Viruses
• Viruses
– Are very small infectious particles consisting of
nucleic acid enclosed in a protein coat and, in
some cases, a membranous envelope

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Viral Genomes
• Viral genomes may consist of
– Double- or single-stranded DNA
– Double- or single-stranded RNA

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Capsids and Envelopes
• A capsid
– Is the protein shell that encloses the viral genome
– Can have various structures
Capsomere
RNA
of capsid
Capsomere DNA

Glycoprotein
18  250 mm 70–90 nm (diameter)

20 nm 50 nm
Figure 18.4a, b (a) Tobacco mosaic virus (b) Adenoviruses
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• Some viruses have envelopes
– Which are membranous coverings derived
from the membrane of the host cell
Membranous
envelope

Capsid
RNA

Glycoprotein
80–200 nm (diameter)

50 nm
Figure 18.4c (c) Influenza viruses
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• Bacteriophages, also called phages
– Have the most complex capsids found among
viruses
Head
Tail DNA
sheath

Tail
fiber

80  225 nm

50 nm
Figure 18.4d (d) Bacteriophage T4
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General Features of Viral Reproductive Cycles
• Viruses are obligate intracellular parasites
– They can reproduce only within a host cell

• Each virus has a host range


– A limited number of host cells that it can infect

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• Viruses use enzymes, ribosomes, and small
molecules of host cells
– To synthesize progeny viruses
VIRUS
Entry into cell and DNA
uncoating of DNA Capsid

Transcription
Replication
HOST CELL

Viral DNA

mRNA

Viral DNA
Capsid
proteins

Self-assembly of
new
virus particles and
their exit from cell
Figure 18.5
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Reproductive Cycles of Phages
• Phages
– Are the best understood of all viruses
– Go through two alternative reproductive
mechanisms: the lytic cycle and the lysogenic
cycle

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The Lytic Cycle
• The lytic cycle
– Is a phage reproductive cycle that culminates
in the death of the host
– Produces new phages and digests the host’s
cell wall, releasing the progeny viruses

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• The lytic cycle of phage T4, a virulent phage
1 Attachment. The T4 phage uses
its tail fibers to bind to specific 2 Entry of phage DNA
receptor sites on the outer and degradation of host DNA.
surface of an E. coli cell. The sheath of the tail contracts,
injecting the phage DNA into
5 Release. The phage directs production the cell and leaving an empty
of an enzyme that damages the bacterial capsid outside. The cell’s
cell wall, allowing fluid to enter. The cell DNA is hydrolyzed.
swells and finally bursts, releasing 100
to 200 phage particles.

Phage assembly

4 Assembly. Three separate sets of proteins 3 Synthesis of viral genomes


self-assemble to form phage heads, tails, and proteins. The phage DNA
and tail fibers. The phage genome is directs production of phage
packaged inside the capsid as the head forms. proteins and copies of the phage
genome by host enzymes, using
Figure 18.6 Head Tails Tail fibers
components within the cell.
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The Lysogenic Cycle
• The lysogenic cycle
– Replicates the phage genome without
destroying the host

• Temperate phages
– Are capable of using both the lytic and
lysogenic cycles of reproduction

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• The lytic and lysogenic cycles of phage , a
temperate phage
Phage
DNA
The phage attaches to a
host cell and injects its DNA.
Many cell divisions
Phage DNA produce a large
circularizes population of bacteria
Phage infected with the
Occasionally, a prophage prophage.
Bacterial exits the bacterial chromosome,
chromosome initiating a lytic cycle.

Lytic cycle Lysogenic cycle


Certain factors The bacterium reproduces
The cell lyses, releasing phages. determine whether normally, copying the prophage
Lytic cycle Lysogenic cycle Prophage and transmitting it to daughter cells.
or
is induced is entered

New phage DNA and Phage DNA integrates into


proteins are synthesized the bacterial chromosome,
Figure 18.7 and assembled into phages. becoming a prophage.

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Reproductive Cycles of Animal Viruses
• The nature of the genome
– Is the basis for the common classification of
animal viruses

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• Classes of animal viruses

Table 18.1
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Viral Envelopes
• Many animal viruses
– Have a membranous envelope

• Viral glycoproteins on the envelope


– Bind to specific receptor molecules on the
surface of a host cell

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• The reproductive cycle of an enveloped RNA virus
1 Glycoproteins on the viral envelope
Capsid
bind to specific receptor molecules
(not shown) on the host cell,
RNA
promoting viral entry into the cell.

Envelope (with
2 Capsid and viral genome
glycoproteins)
enter cell

HOST CELL
3 The viral genome (red)
functions as a template for
Viral genome (RNA) synthesis of complementary
RNA strands (pink) by a viral
Template enzyme.

5 Complementary RNA mRNA


strands also function as mRNA, Capsid
which is translated into both proteins
capsid proteins (in the cytosol) 4 New copies of viral
and glycoproteins for the viral ER genome RNA are made
Glyco- Copy of
envelope (in the ER). using complementary RNA
proteins genome (RNA)
strands as templates.

6 Vesicles transport
envelope glycoproteins to
the plasma membrane.
8 New virus
7 A capsid assembles
around each viral
Figure 18.8 genome molecule.
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RNA as Viral Genetic Material
• The broadest variety of RNA genomes
– Is found among the viruses that infect animals

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• Retroviruses, such as HIV, use the enzyme
reverse transcriptase
– To copy their RNA genome into DNA, which
can then be integrated into the host genome
as a provirus
Glycoprotein
Viral envelope

Capsid

Reverse RNA
transcriptase (two identical
strands)
Figure 18.9
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• The reproductive cycle of HIV, a retrovirus 1 The virus fuses with the
HIV Membrane of cell’s plasma membrane.
white blood cell The capsid proteins are 2 Reverse transcriptase
removed, releasing the catalyzes the synthesis of a
viral proteins and RNA. DNA strand complementary
to the viral RNA.

HOST CELL

Reverse
3 Reverse transcriptase
catalyzes the synthesis of
transcriptase
a second DNA strand
Viral RNA
complementary to the first.

RNA-DNA
hybrid 4 The double-stranded
0.25 µm DNA is incorporated
as a provirus into the
HIV entering a cell
DNA cell’s DNA.

NUCLEUS
Chromosomal Provirus
5 Proviral genes are
DNA transcribed into RNA
RNA genome molecules, which serve as
for the next genomes for the next viral
viral generation generation and as mRNAs
mRNA for translation into viral
proteins.

6 The viral proteins include


capsid proteins and reverse
transcriptase (made in the cytosol)
and envelope glycoproteins
(made in the ER).

8 Capsids are 7 Vesicles transport the


assembled around glycoproteins from the ER to
viral genomes and the cell’s plasma membrane.
9 New viruses bud reverse transcriptase
Figure 18.10 New HIV leaving a cell off from the host cell. molecules.

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Evolution of Viruses
• Viruses do not really fit our definition of living
organisms
• Since viruses can reproduce only within cells
– They probably evolved after the first cells
appeared, perhaps packaged as fragments of
cellular nucleic acid

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• Concept 18.2: Viruses, viroids, and prions are
formidable pathogens in animals and plants
• Diseases caused by viral infections
– Affect humans, agricultural crops, and livestock
worldwide

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Viral Diseases in Animals
• Viruses may damage or kill cells
– By causing the release of hydrolytic enzymes
from lysosomes

• Some viruses cause infected cells


– To produce toxins that lead to disease
symptoms

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• Vaccines
– Are harmless derivatives of pathogenic
microbes that stimulate the immune system to
mount defenses against the actual pathogen
– Can prevent certain viral illnesses

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Emerging Viruses
• Emerging viruses
– Are those that appear suddenly or suddenly
come to the attention of medical scientists

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• Severe acute respiratory syndrome (SARS)
– Recently appeared in China

(a) Young ballet students in Hong Kong (b) The SARS-causing agent is a coronavirus
wear face masks to protect themselves like this one (colorized TEM), so named for the
from the virus causing SARS. “corona” of glycoprotein spikes protruding from
Figure 18.11 A, B the envelope.
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• Outbreaks of “new” viral diseases in humans
– Are usually caused by existing viruses that
expand their host territory

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Viral Diseases in Plants
• More than 2,000 types of viral diseases of
plants are known
• Common symptoms of viral infection include
– Spots on leaves and fruits, stunted growth, and
damaged flowers or roots

Figure 18.12
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• Plant viruses spread disease in two major
modes
– Horizontal transmission, entering through
damaged cell walls
– Vertical transmission, inheriting the virus from
a parent

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Viroids and Prions: The Simplest Infectious Agents
• Viroids
– Are circular RNA molecules that infect plants
and disrupt their growth

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• Prions
– Are slow-acting, virtually indestructible
infectious proteins that cause brain diseases in
mammals
– Propagate by converting normal proteins into
the prion version
Prion Original
prion

Many prions

Normal
New
protein
prion

Figure 18.13
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• Concept 18.3: Rapid reproduction, mutation,
and genetic recombination contribute to the
genetic diversity of bacteria
• Bacteria allow researchers
– To investigate molecular genetics in the
simplest true organisms

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The Bacterial Genome and Its Replication
• The bacterial chromosome
– Is usually a circular DNA molecule with few
associated proteins

• In addition to the chromosome


– Many bacteria have plasmids, smaller circular
DNA molecules that can replicate
independently of the bacterial chromosome

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• Bacterial cells divide by binary fission
– Which is preceded by replication of the
bacterial chromosome
Replication
fork

Origin of
replication

Termination
of replication

Figure 18.14
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Mutation and Genetic Recombination as Sources
of Genetic Variation

• Since bacteria can reproduce rapidly


– New mutations can quickly increase a
population’s genetic diversity

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• Further genetic diversity
– Can arise by recombination of the DNA from
two different bacterial cells
EXPERIMENT Researchers had two mutant strains, one that could make arginine but not
tryptophan trp–) and one that could make tryptophan but not arginine (arg trp+). Each
(arg+
mutant strain and a mixture of both strains were grown in a liquid medium containing all the
required amino acids. Samples from each liquid culture were spread on plates containing a
solution of glucose and inorganic salts (minimal medium), solidified with agar.
Mixture

Mutant Mutant
strain strain
arg+ trp– arg trp+

RESULTS Only the samples from the mixed culture, contained cells that gave rise to colonies on
Figure 18.15 minimal medium, which lacks amino acids.
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Mixture

Mutant Mutant
strain strain
arg+ trp– arg– trp+

No No
colonies colonies
(control) Colonies (control)
grew

CONCLUSION Because only cells that can make both arginine and tryptophan (arg+ trp+ cells) can grow into
colonies on minimal medium, the lack of colonies on the two control plates showed that no further mutations had
occurred restoring this ability to cells of the mutant strains. Thus, each cell from the mixture that formed a colony on the
minimal medium must have acquired one or more genes from a cell of the other strain by genetic recombination.

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Mechanisms of Gene Transfer and Genetic
Recombination in Bacteria

• Three processes bring bacterial DNA from


different individuals together
– Transformation
– Transduction
– Conjugation

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Transformation
• Transformation
– Is the alteration of a bacterial cell’s genotype
and phenotype by the uptake of naked, foreign
DNA from the surrounding environment

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Transduction
• In the process known as transduction
– Phages carry bacterial genes from one host
cell to another Phage DNA

1 Phage infects bacterial cell that has alleles A+ and B+ A+ B+

2 Host DNA (brown) is fragmented, and phage DNA A+ B+


and proteins are made. This is the donor cell.

Donor
cell

3 A bacterial DNA fragment (in this case a fragment with


the A+ allele) may be packaged in a phage capsid.

A+
Crossing
over
4 Phage with the A+ allele from the donor cell infects
a recipient A–B– cell, and crossing over (recombination) A+
between donor DNA (brown) and recipient DNA
(green) occurs at two places (dotted lines). A– B–

Recipient
cell

5 The genotype of the resulting recombinant cell (A+B–)


differs from the genotypes of both the donor (A+B+) and A+ B–
Figure 18.16 the recipient (A–B–). Recombinant cell

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Conjugation and Plasmids
• Conjugation
– Is the direct transfer of genetic material between
bacterial cells that are temporarily joined

Figure 18.17 Sex pilus 1 m


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The F Plasmid and Conjugation
• Cells containing the F plasmid, designated F+
cells
– Function as DNA donors during conjugation
– Transfer plasmid DNA to an F recipient cell

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• Conjugation and transfer of an F plasmid from
an F+ donor to an F recipient

F Plasmid Bacterial chromosome

F+ cell
F+ cell
Mating
bridge

Bacterial F+ cell
F– cell
chromosome

1 A cell carrying an F plasmid 2 A single strand of the 3 DNA replication occurs in 4 The plasmid in the
(an F+ cell) can form a F plasmid breaks at a both donor and recipient recipient cell
mating bridge with an F– cell specific point (tip of blue cells, using the single circularizes. Transfer
and transfer its F plasmid. arrowhead) and begins to parental strands of the and replication result
move into the recipient cell. F plasmid as templates in a compete F plasmid (a) Conjugation and transfer of an
As transfer continues, the to synthesize complementary in each cell. Thus, both
donor plasmid rotates strands. cells are now F+. F plasmid from an F+ donor to
(red arrow). an F– recipient
Figure 18.18a

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• Chromosomal genes can be transferred during
conjugation
– When the donor cell’s F factor is integrated into the
chromosome
• A cell with the F factor built into its chromosome
– Is called an Hfr cell
• The F factor of an Hfr cell
– Brings some chromosomal DNA along with it when it
is transferred to an F– cell

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• Conjugation and transfer of part of the
bacterial chromosome from an Hfr donor
to an F– recipient, resulting in recombination
F+ cell Hfr cell
F factor
1 The circular F plasmid in an F + cell
can be integrated into the circular 2 The resulting cell is called an Hfr cell
chromosome by a single crossover (for High frequency of recombination).
event (dotted line).

B+ C+ C+ D+
A+ D+ A+
Hfr cell B+ A+ D+ C+
B+ D + C+ B+
A+

A+

B+
C– C– C– D – C–
F– cell B–
A–
D– B–
A–
D– A+ B–
A– A+
B+ B–
A–
D–

3 Since an Hfr cell has all 4 A single strand of the F factor 5 The location and orientation 6 The mating bridge
the F-factor genes, it can breaks and begins to move of the F factor in the donor usually breaks well
form a mating bridge with through the bridge. DNA chromosome determine before the entire
an F– cell and transfer DNA. replication occurs in both donor the sequence of gene transfer chromosome and
and recipient cells, resulting in during conjugation. In this the rest of the
double-stranded DNA example, the transfer sequence F factor are transferred.
for four genes is A-B-C-D.

Temporary
B+ C– B– C– Recombinant F–
partial B–
A–
D– B+
A–
D–
A+ A+
diploid bacterium

7 Two crossovers can result 8 The piece of DNA ending up outside the
in the exchange of similar bacterial chromosome will eventually be
(homologous) genes between degraded by the cell’s enzymes. The recipient
the transferred chromosome fragment cell now contains a new combination of genes (b) Conjugation and transfer of part
(brown) and the recipient cell’s but no F factor; it is a recombinant F – cell.
chromosome (green). of the bacterial chromosome from
an Hfr donor to an F– recipient,
Figure 18.18b resulting in recombination

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R plasmids and Antibiotic Resistance
• R plasmids
– Confer resistance to various antibiotics

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Transposition of Genetic Elements
• Transposable elements
– Can move around within a cell’s genome
– Are often called “jumping genes”
– Contribute to genetic shuffling in bacteria

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Insertion Sequences
• An insertion sequence contains a single gene
for transposase
– An enzyme that catalyzes movement of the
insertion sequence from one site to another
within the genome
Insertion sequence

3 A T C C G G T… A C C G G A T… 3
5 TAG G C CA… TG G C CTA… 5
Inverted Transposase gene Inverted
repeat repeat
(a) Insertion sequences, the simplest transposable elements in bacteria, contain a single gene that
encodes transposase, which catalyzes movement within the genome. The inverted repeats are
backward, upside-down versions of each other; only a portion is shown. The inverted repeat
sequence varies from one type of insertion sequence to another.

Figure 18.19a

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Transposons
• Bacterial transposons
– Also move about within the bacterial genome
– Have additional genes, such as those for
antibiotic resistance
Transposon

Insertion Antibiotic Insertion


sequence resistance gene sequence

5 5
3 3

Inverted repeats Transposase gene


(b) Transposons contain one or more genes in addition to the transposase gene. In the transposon
shown here, a gene for resistance to an antibiotic is located between twin insertion sequences.
The gene for antibiotic resistance is carried along as part of the transposon when the transposon
is inserted at a new site in the genome.

Figure 18.19b

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• Concept 18.4: Individual bacteria respond to
environmental change by regulating their gene
expression
• E. coli, a type of bacteria that lives in the
human colon
– Can tune its metabolism to the changing
environment and food sources

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• This metabolic control occurs on two levels
– Adjusting the activity of metabolic enzymes
already present
– Regulating the genes encoding the metabolic
enzymes (a) Regulation of enzyme
activity
Precursor
(b) Regulation of enzyme
production

Feedback
inhibition
Enzyme 1 Gene 1

Enzyme 2 Gene 2 Regulation


of gene
expression
Enzyme 3 Gene 3


Enzyme 4 Gene 4


Enzyme 5 Gene 5

Tryptophan
Figure 18.20a, b
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Operons: The Basic Concept
• In bacteria, genes are often clustered into
operons, composed of
– An operator, an “on-off” switch
– A promoter
– Genes for metabolic enzymes

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• An operon
– Is usually turned “on”
– Can be switched off by a protein called a
repressor

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• The trp operon: regulated synthesis of
repressible enzymes
trp operon
Promoter
Promoter
Genes of operon
DNA trpR trpE trpD trpC trpB trpA
Operator
Regulatory RNA Start codon Stop codon
gene 3 polymerase
mRNA 5
mRNA
5
E D C B A
Protein Inactive
repressor Polypeptides that make up
enzymes for tryptophan synthesis

(a) Tryptophan absent, repressor inactive, operon on. RNA polymerase attaches to the DNA at the
promoter and transcribes the operon’s genes.

Figure 18.21a
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DNA

No RNA made

mRNA

Active
Protein repressor

Tryptophan
(corepressor)
(b) Tryptophan present, repressor active, operon off. As tryptophan
accumulates, it inhibits its own production by activating the repressor protein.
Figure 18.21b
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Repressible and Inducible Operons: Two Types of
Negative Gene Regulation
• In a repressible operon
– Binding of a specific repressor protein to the
operator shuts off transcription

• In an inducible operon
– Binding of an inducer to an innately inactive
repressor inactivates the repressor and turns
on transcription

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• The lac operon: regulated synthesis of
inducible enzymes Promoter
Regulatory
Operator
gene
DNA lacl lacZ

No
RNA
made
3
RNA
mRNA polymerase
5

Active
Protein repressor

(a) Lactose absent, repressor active, operon off. The lac repressor is innately active, and in
the absence of lactose it switches off the operon by binding to the operator.
Figure 18.22a
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lac operon

DNA lacl lacz lacY lacA

RNA
polymerase
3
mRNA mRNA 5'
mRNA 5
5

Protein -Galactosidase Permease Transacetylase

Allolactose Inactive
(inducer) repressor

(b) Lactose present, repressor inactive, operon on. Allolactose, an isomer of lactose, derepresses
the operon by inactivating the repressor. In this way, the enzymes for lactose utilization are induced.
Figure 18.22b

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• Inducible enzymes
– Usually function in catabolic pathways

• Repressible enzymes
– Usually function in anabolic pathways

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• Regulation of both the trp and lac operons
– Involves the negative control of genes,
because the operons are switched off by the
active form of the repressor protein

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Positive Gene Regulation
• Some operons are also subject to positive
control
– Via a stimulatory activator protein, such as
catabolite activator protein (CAP)

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• In E. coli, when glucose, a preferred food
source, is scarce
– The lac operon is activated by the binding of a
regulatory protein, catabolite activator protein
(CAP) Promoter

DNA lacl lacZ

CAP-binding site RNA Operator


polymerase
can bind
Active
cAMP and transcribe
CAP

Inactive lac
Inactive repressor
CAP

(a) Lactose present, glucose scarce (cAMP level high): abundant lac mRNA synthesized.
If glucose is scarce, the high level of cAMP activates CAP, and the lac operon produces
Figure 18.23a large amounts of mRNA for the lactose pathway.
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• When glucose levels in an E. coli cell increase
– CAP detaches from the lac operon, turning it
off
Promoter
DNA
lacl lacZ

CAP-binding site Operator


RNA
polymerase
can’t bind

Inactive
CAP Inactive lac
repressor

(b) Lactose present, glucose present (cAMP level low): little lac mRNA synthesized.
Figure 18.23b When glucose is present, cAMP is scarce, and CAP is unable to stimulate transcription.

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