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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

Figure 18.1
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0.5 m

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

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

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

Figure 18.2

0.25 m

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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
of capsid

RNA
Capsomere

DNA

Glycoprotein
7090 nm (diameter)
18 250 mm

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
80200 nm (diameter)

Figure 18.4c

50 nm
(c) Influenza viruses

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Bacteriophages, also called phages


Have the most complex capsids found among
viruses
Head

Tail
sheath

DNA

Tail
fiber

80 225 nm

Figure 18.4d
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50 nm
(d) Bacteriophage T4

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
Entry into cell and
uncoating of DNA

DNA
Capsid

VIRUS

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 hosts
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

2 Entry of phage DNA

its tail fibers to bind to specific


receptor sites on the outer
surface of an E. coli cell.

and degradation of host DNA.


The sheath of the tail contracts,
injecting the phage DNA into
the cell and leaving an empty
capsid outside. The cells
DNA is hydrolyzed.

5 Release. The phage directs production


of an enzyme that damages the bacterial
cell wall, allowing fluid to enter. The cell
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 tail fibers. The phage genome is
packaged inside the capsid as the head forms.
Figure 18.6

Head

Tails

Tail fibers

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and proteins. The phage DNA


directs production of phage
proteins and copies of the phage
genome by host enzymes, using
components within the cell.

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.
Phage DNA
circularizes
Phage
Occasionally, a prophage
exits the bacterial chromosome,
initiating a lytic cycle.

Bacterial
chromosome

Lytic cycle
The cell lyses, releasing phages.

Lysogenic cycle
Certain factors
determine whether
Lytic cycle
is induced

Figure 18.7

Many cell divisions


produce a large
population of bacteria
infected with the
prophage.

or

New phage DNA and


proteins are synthesized
and assembled into phages.

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Lysogenic cycle
is entered

Prophage

The bacterium reproduces


normally, copying the prophage
and transmitting it to daughter cells.

Phage DNA integrates into


the bacterial chromosome,
becoming a prophage.

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,
promoting viral entry into the cell.

RNA
Envelope (with
glycoproteins)

2 Capsid and viral genome


enter cell

HOST CELL
Viral genome (RNA)
Template
5 Complementary RNA
strands also function as mRNA,
which is translated into both
capsid proteins (in the cytosol)
and glycoproteins for the viral
envelope (in the ER).

3 The viral genome (red)


functions as a template for
synthesis of complementary
RNA strands (pink) by a viral
enzyme.

mRNA
Capsid
proteins
ER

Glycoproteins

Copy of
genome (RNA)

4 New copies of viral


genome RNA are made
using complementary RNA
strands as templates.

6 Vesicles transport
envelope glycoproteins to
the plasma membrane.
8 New virus
7 A capsid assembles

Figure 18.8

around each viral


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
transcriptase

Figure 18.9
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RNA
(two identical
strands)

The reproductive cycle of HIV, a retrovirus


HIV

Membrane of
white blood cell

1 The virus fuses with the


cells plasma membrane.
The capsid proteins are
removed, releasing the
viral proteins and RNA.

2 Reverse transcriptase
catalyzes the synthesis of a
DNA strand complementary
to the viral RNA.
HOST CELL

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

Reverse
transcriptase
Viral RNA

0.25 m
HIV entering a cell

RNA-DNA
hybrid

4 The double-stranded
DNA is incorporated
as a provirus into the
cells DNA.

DNA

NUCLEUS
Chromosomal
DNA
RNA genome
for the next
viral generation

Provirus

mRNA

5 Proviral genes are


transcribed into RNA
molecules, which serve as
genomes for the next viral
generation and as mRNAs
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).

Figure 18.10

New HIV leaving a cell

9 New viruses bud


off from the host cell.

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8 Capsids are
assembled around
viral genomes and
reverse transcriptase
molecules.

7 Vesicles transport the


glycoproteins from the ER to
the cells plasma membrane.

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
like this one (colorized TEM), so named for the
wear face masks to protect themselves
corona of glycoprotein spikes protruding from
from the virus causing SARS.
the envelope.
Figure 18.11 A, B
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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
protein

New
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
populations 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 (arg+ trp) and one that could make tryptophan but not arginine (arg trp+). Each
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
strain
arg+ trp

Figure 18.15

Mutant
strain
arg trp+

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

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Mixture

Mutant
strain
arg+ trp

Mutant
strain
arg trp+

No
colonies
(control)

CONCLUSION

Colonies
grew

No
colonies
(control)

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 cells 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

A+ B+

1 Phage infects bacterial cell that has alleles A+ and 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+
4 Phage with the A+ allele from the donor cell infects

a recipient AB cell, and crossing over (recombination)


between donor DNA (brown) and recipient DNA
(green) occurs at two places (dotted lines).

Crossing
over
A+
A B
Recipient
cell

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

Figure 18.16

differs from the genotypes of both the donor (A+B+) and


the recipient (AB).

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A+ B
Recombinant cell

Conjugation and Plasmids


Conjugation
Is the direct transfer of genetic material between
bacterial cells that are temporarily joined

Figure 18.17
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Sex pilus

1 m

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
F cell

Bacterial
chromosome

1 A cell carrying an F plasmid


(an F+ cell) can form a
mating bridge with an F cell
and transfer its F plasmid.

2 A single strand of the


F plasmid breaks at a
specific point (tip of blue
arrowhead) and begins to
move into the recipient cell.
As transfer continues, the
donor plasmid rotates
(red arrow).

F+ cell
3 DNA replication occurs in 4
both donor and recipient
cells, using the single
parental strands of the
F plasmid as templates
to synthesize complementary
strands.

Figure 18.18a

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The plasmid in the


recipient cell
circularizes. Transfer
and replication result
in a compete F plasmid
in each cell. Thus, both
cells are now F+.

(a) Conjugation and transfer of an


F plasmid from an F+ donor to
an F recipient

Chromosomal genes can be transferred during


conjugation
When the donor cells 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
Hfr cell

F+ cell
1

Hfr cell

A+

F factor

The circular F plasmid in an F+ cell


can be integrated into the circular
chromosome by a single crossover
event (dotted line).

B+

C+

The resulting cell is called an Hfr cell


(for High frequency of recombination).

D+

C+
B+

D+

D+ C+

A+

B+

A+

D+ C+

B+

A+

A+

F cell
3

C
A

B+
D

Since an Hfr cell has all


the F-factor genes, it can
form a mating bridge with
an F cell and transfer DNA.

B+
A+

C
A

A+

4 A single strand of the F factor


breaks and begins to move
through the bridge. DNA
replication occurs in both donor
and recipient cells, resulting in
double-stranded DNA

Temporary
partial
diploid
7

C D
A+

5 The location and orientation


of the F factor in the donor
chromosome determine
the sequence of gene transfer
during conjugation. In this
example, the transfer sequence
for four genes is A-B-C-D.

Two crossovers can result


in the exchange of similar
(homologous) genes between
the transferred chromosome fragment
(brown) and the recipient cells
chromosome (green).

Figure 18.18b
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B
A+

B+

B+

C
A

C
A

The mating bridge


usually breaks well
before the entire
chromosome and
the rest of the
F factor are transferred.

Recombinant F
bacterium

8 The piece of DNA ending up outside the


bacterial chromosome will eventually be
degraded by the cells enzymes. The recipient
cell now contains a new combination of genes
but no F factor; it is a recombinant F cell.

(b) Conjugation and transfer of part


of the bacterial chromosome from
an Hfr donor to an F recipient,
resulting in recombination

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 cells 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

TAG G C CA

TG G C C TA

Transposase gene
Inverted
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
sequence

Antibiotic
resistance gene

Insertion
sequence

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

Feedback
inhibition

(b) Regulation of enzyme


production

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

DNA

Promoter
Genes of operon
trpD
trpC
trpE

trpR

trpB

trpA

Operator
Regulatory
gene
mRNA
5

RNA
polymerase

Start codon

Stop codon

mRNA 5
E

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 operons genes.
Figure 18.21a
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DNA
No RNA made

mRNA

Protein

Active
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
gene

DNA

Operator

lacl

lacZ

3
mRNA

Protein

No
RNA
made
RNA
polymerase

Active
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

3
mRNA

lacA

RNA
polymerase
mRNA 5'
5
mRNA

-Galactosidase

Protein
Allolactose
(inducer)

lacY

Permease

Transacetylase

Inactive
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
Promoter
(CAP)
DNA

lacl

lacZ

CAP-binding site
cAMP

Inactive
CAP

RNA
Operator
polymerase
can bind
Active
and transcribe
CAP

Inactive lac
repressor

(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
cant bind

Inactive
CAP

Inactive lac
repressor

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

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