CAMPBELL BIOLOGY IN FOCUS

Urry • Cain • Wasserman • Minorsky • Jackson • Reece

14
Gene Expression:
From Gene to Protein

Lecture Presentations by
Kathleen Fitzpatrick and Nicole Tunbridge

© 2014 Pearson Education, Inc.

Overview: The Flow of Genetic Information

 The information content of genes is in the form of

specific sequences of nucleotides in DNA
 The DNA inherited by an organism leads to specific
traits by dictating the synthesis of proteins
 Proteins are the links between genotype and
phenotype
 Gene expression, the process by which DNA
directs protein synthesis, includes two stages:
transcription and translation

© 2014 Pearson Education, Inc.

Figure 14.1

© 2014 Pearson Education,

Concept 14.1: Genes specify proteins via
transcription and translation
 How was the fundamental relationship between
genes and proteins discovered?

© 2014 Pearson Education,

Evidence from the Study of Metabolic Defects

 In 1902, British physician Archibald Garrod first

suggested that genes dictate phenotypes through
enzymes that catalyze specific chemical reactions
 He thought symptoms of an inherited disease reflect
an inability to synthesize a certain enzyme
 Cells synthesize and degrade molecules in a series
of steps, a metabolic pathway

© 2014 Pearson Education,

Nutritional Mutants in Neurospora: Scientific
Inquiry

© 2014 Pearson Education,

 Figure

Control: Wild-type
2 Cells subjected to X-rays. Growth cells in minimal
Neurospora medium
cells

4 Surviving cells
No growth tested for inability
to grow on
minimal medium.

3 Each
1 Individual Neurospora cells placed on surviving
complete cell forms
growth a colony of genetically identical cell
medium.
5 Mutant cells placed
Growth in a series of vials,
each containing
minimal medium
plus one additional
nutrient.

© 2014 Pearson Education,

 Figure

2 Cells subjected
Neurospora to X-rays.
cells

Individual Neurospora
1 3 Each surviving
cells placed on
cell forms a
complete growth
colony of
medium.
genetically
identical cells.

© 2014 Pearson Education,

 Figure

Control: Wild-type
Growth cells in minimal
medium

4 Surviving cells
No tested for inability
growth to grow on
minimal medium.

5 Mutant cells placed

Growth in a series of vials,
each containing
minimal medium
plus one additional
nutrient.
© 2014 Pearson Education,
 The researchers amassed a valuable collection of
Neurospora mutant strains, catalogued by their
defects
 For example, one set of mutants all required
arginine for growth
 It was determined that different classes of these
mutants were blocked at a different step in the
biochemical pathway for arginine biosynthesis

© 2014 Pearson Education,

Figure 14.3

Gene A Gene B Gene C

Enzyme Enzyme Enzyme

A B C
Precursor Ornithine Citrulline Arginine

© 2014 Pearson Education,

The Products of Gene Expression: A Developing
Story

© 2014 Pearson Education,

Basic Principles of Transcription and Translation

© 2014 Pearson Education,

© 2014 Pearson Education,
© 2014 Pearson Education,
 Figure

Nuclear
envelope

TRANSCRIPTION DNA

RNA PROCESSING Pre-mRNA

mRNA
DNA
TRANSCRIPTION

mRNA
Ribosome TRANSLATION Ribosome
TRANSLATION

Polypeptide Polypeptide

(a) Bacterial cell (b) Eukaryotic cell

© 2014 Pearson Education,

 Figure 14.4a-

DNA
TRANSCRIPTION

mRNA

(a) Bacterial cell

© 2014 Pearson Education,

 Figure 14.4a-

DNA
TRANSCRIPTION

mRNA
Ribosome
TRANSLATION

Polypeptide

(a) Bacterial cell

© 2014 Pearson Education,

 Figure 14.4b-
Nuclear
envelope

TRANSCRIPTION DNA

Pre-mRNA

(b) Eukaryotic
© 2014 Pearson Education,
 Figure 14.4b-
Nuclear
envelope

TRANSCRIPTION DNA

RNA PROCESSING Pre-mRNA

mRNA

(b) Eukaryotic
© 2014 Pearson Education,
 Figure 14.4b-
Nuclear
envelope

TRANSCRIPTION DNA

RNA PROCESSING Pre-mRNA

mRNA

TRANSLATION
Ribosome

Polypeptide

(b) Eukaryotic
© 2014 Pearson Education,
 A primary transcript is the initial RNA transcript
from any gene prior to processing
 The central dogma is the concept that cells are
governed by a cellular chain of command

© 2014 Pearson Education,

Figure 14.UN01

DNA RNA Protein

© 2014 Pearson Education,

The Genetic Code

 How are the instructions for assembling amino acids

into proteins encoded into DNA?
 There are 20 amino acids, but there are only four
nucleotide bases in DNA
 How many nucleotides correspond to an amino acid?

© 2014 Pearson Education,

Codons: Triplets of Nucleotides

© 2014 Pearson Education,

Figure 14.5

DNA
template
3 5
strand
A C C A A A C C G A G T

T G G T T T G G C T C A
5 3

TRANSCRIPTION

U G G U U U G G C U C A
mRNA 5 3
Codon
TRANSLATION

Protein Trp Phe Gly Ser

© 2014 Pearson Education,

 Amino acid

© 2014 Pearson Education,

© 2014 Pearson Education,
© 2014 Pearson Education,
Cracking the Code

© 2014 Pearson Education,

© 2014 Pearson Education,
Figure 14.6
Second mRNA base
U C A G
UUU
UCU UAU UGU U
UUC Phe Tyr Cys
U UCC UAC UGC C
UUA Ser
UCA UAA Stop UGA Stop A
UUG Leu
UCG UAG Stop UGG Trp G
CUU
CCU CAU CGU U
First mRNA base (5 end of

His

Third mRNA base (3 end of

CUC
C CCC CAC CGC C
CUA Leu Pro Arg
CCA CAA CGA A
CUG Gln
CCG CAG CGG G
AUU ACU AAU AGU U
Asn Ser
AUC IIe ACC AAC AGC C
A Thr
AUA ACA AAA AGA A
Lys Arg
Met or
AUG start ACG AAG AGG G

GUU GCU GAU GGU U

Asp
GUC GCC GAC GGC C
G Val Ala Gly
GUA GCA GAA GGA A
Glu
GUG GCG GAG GGG G
© 2014 Pearson Education,
Evolution of the Genetic Code

© 2014 Pearson Education,

 Figure

(a) Tobacco plant (b) Pig expressing a jellyfish

expressing a firefly gene gene

© 2014 Pearson Education,

 Figure

(a) Tobacco plant expressing

a firefly gene

© 2014 Pearson Education,

 Figure

(b) Pig expressing a jellyfish

gene

© 2014 Pearson Education,

Concept 14.2: Transcription is the DNA-directed
synthesis of RNA: a closer look
 Transcription is the first stage of gene expression

© 2014 Pearson Education,

Molecular Components of Transcription

Animation: Transcription Introduction

© 2014 Pearson Education,

 Figure 14.8-
Promote Transcription
5 3
3 5
Start point
Initiation RNA polymerase
1
5 3
3 5
Unwound RNA Template strand of DNA
DNA transcript

© 2014 Pearson Education,

 Figure 14.8-
Promote Transcription
5 3
3 5
Start point
Initiation RNA polymerase
1
5 3
3 5
Unwound RNA Template strand of DNA
DNA transcript
2 Elongation
Rewound
DNA
5 3
3 3 5
5
RNA Direction of
transcript transcription
(“downstream”)

© 2014 Pearson Education,

 Figure 14.8-
Promote Transcription
5 3
3 5
Start point
Initiation RNA polymerase
1
5 3
3 5
Unwound RNA Template strand of DNA
DNA transcript
2 Elongation
Rewound
DNA 3
5 3 5
3
5
Direction of
RNA transcription
transcript (“downstream”)
3 Termination
3
5 5
3 3
5
Completed RNA transcript
© 2014 Pearson Education,
© 2014 Pearson Education,
Synthesis of an RNA Transcript

 The three stages of transcription

 Initiation
 Elongation
 Termination

© 2014 Pearson Education,

RNA Polymerase Binding and Initiation of
Transcription

© 2014 Pearson Education,

© 2014 Pearson Education,
 Figure

TRANSCRIPTION DNA

RNA PROCESSING Pre-mRNA

mRNA

TRANSLATION Ribosome

Polypeptide
© 2014 Pearson Education,
 Figure
Promoter
Nontemplate strand
DNA
5 TATAAAA 3
3 ATAT T T T 5 1 A eukaryotic
TATA box Start point Template promoter
Transcription strand
factors

5 3
3 5 2 Several transcription
factors bind to DNA.

RNA polymerase II Transcription

factors 3 Transcription
initiation
complex forms.
5 3
5
3 5
3

RNA transcript
© 2014 Pearson Education,
 Figure

Transcription initiation complex

© 2014 Pearson Education,

Elongation of the RNA Strand

 As RNA polymerase moves along the DNA, it

untwists the double helix, 10 to 20 bases at a time
 Transcription progresses at a rate of 40 nucleotides
per second in eukaryotes
 A gene can be transcribed simultaneously by
several RNA polymerases

© 2014 Pearson Education,

Figure 14.10

Nontemplate
strand of DNA
RNA nucleotides
RNA
polymerase

A T C
3 C A A
5
3 end

A U C C A
5 3
T A G
G T T

5 Direction of transcription
Template
strand of DNA
Newly made
RNA
© 2014 Pearson Education,
Termination of Transcription

© 2014 Pearson Education,

Concept 14.3: Eukaryotic cells modify RNA after
transcription

© 2014 Pearson Education,

Alteration of mRNA Ends

© 2014 Pearson Education,

 Figure

TRANSCRIPTION DNA

RNA PROCESSING Pre-mRNA

mRNA

TRANSLATION Ribosome

Polypeptide
© 2014 Pearson Education,
 Figure

A modified guanine 50–250 adenine

nucleotide added to nucleotides added
the 5 end to the 3 end
Protein-coding segment Polyadenylation
5 signal
3
G P P P AAUAAA AAA…AAA

5 Cap 5 Start Stop

UTR 3 UTR Poly-A tail
codon codon

© 2014 Pearson Education,

Split Genes and RNA Splicing

 Most eukaryotic mRNAs have long noncoding

stretches of nucleotides that lie between coding
regions
 The noncoding regions are called intervening
sequences, or introns
 The other regions are called exons and are usually
translated into amino acid sequences
 RNA splicing removes introns and joins exons,
creating an mRNA molecule with a continuous
coding sequence

© 2014 Pearson Education,

Figure 14.12

Pre-mRNA
Intron Intron
5 Cap Poly-A tail
1–30 31–104 105–
Introns cut out and 146
exons spliced together
mRNA
5 Cap Poly-A tail
1–146
5 UTR 3 UTR
Coding
segment

© 2014 Pearson Education,

© 2014 Pearson Education,
Figure 14.13

Spliceosome
Small RNAs

5

Pre-mRNA

Exon 1 Exon 2

Intron

Spliceosome
components
mRNA
5 Cut-out
Exon 1 Exon 2 intron
© 2014 Pearson Education,
Ribozymes

© 2014 Pearson Education,

Concept 14.4: Translation is the RNA-directed
synthesis of a polypeptide: a closer look
 Genetic information flows from mRNA to protein
through the process of translation

© 2014 Pearson Education,

Molecular Components of Translation

 A cell translates an mRNA message into protein with

the help of transfer RNA (tRNA)
 tRNAs transfer amino acids to the growing
polypeptide in a ribosome
 Translation is a complex process in terms of its
biochemistry and mechanics

© 2014 Pearson Education,

 Figure

TRANSCRIPTION DNA

mRNA
Ribosome
TRANSLATION

Polypeptide

© 2014 Pearson Education,

 Figure

Amino acids
Polypeptide

tRNA with
amino acid
attached
Ribosome

Gly

tRNA

Anticodon
A A A
U G G U U U G G C

5 Codons 3
mRNA
© 2014 Pearson Education,
The Structure and Function of Transfer RNA

 Each tRNA can translate a particular mRNA codon

into a given amino acid
 The tRNA contains an amino acid at one end and at
the other end has a nucleotide triplet that can base-
pair with the complementary codon on mRNA

© 2014 Pearson Education,

 A tRNA molecule consists of a single RNA strand
that is only about 80 nucleotides long
 tRNA molecules can base-pair with themselves
 Flattened into one plane, a tRNA molecule looks like
a cloverleaf
 In three dimensions, tRNA is roughly L-shaped,
where one end of the L contains the anticodon that
base-pairs with an mRNA codon

Video: tRNA Model

© 2014 Pearson Education,

Figure 14.15

3
A
Amino C
acid C
attachment A 5
site C G Amino acid
G C attachment site
C G 5
U G
U A 3
A U
U A U
C CA C A G U A G *
* A * C U C
G *
C
G U G U *
C G A G G Hydrogen
* G
*
U
C
*
A
G bonds
*GA C
G
G
Hydrogen
C
U bonds
A
* G
* A
A C
* A A G
U 3 5
A G
A Anticodon Anticodon
Anticodon
(b) Three-dimensional (c) Symbol used
(a) Two-dimensional structure structure in this book

© 2014 Pearson Education,

Figure 14.15a
3
A
Amino acid attachment
C site
CACGCUU
A 5
GCGGAUU
U

U C A
* C A C AG A G *
A * C U C
G C *G
U G U *
C G G A G A G G
* * U C
* G AG *
GU CCAG
* A Hydrogen bonds
*
A
*
A
CU
G

Anticodon
(a) Two-dimensional structure
© 2014 Pearson Education,
Figure 14.15b

Amino acid
attachment site
5
3

Hydrogen
bonds

A A G
3 5
Anticodon Anticodon

(b) Three-dimensional (c) Symbol used

structure in this book

© 2014 Pearson Education,

 Accurate translation requires two steps
 First: a correct match between a tRNA and an amino
acid, done by the enzyme aminoacyl-tRNA
synthetase
 Second: a correct match between the tRNA
anticodon and an mRNA codon
 Flexible pairing at the third base of a codon is
called wobble and allows some tRNAs to bind to
more than one codon

© 2014 Pearson Education,

 Figure 14.16-
1 Amino acid Tyrosine Tyrosyl-tRNA
and tRNA (Tyr) (amino synthetase
enter active acid)
site.

Tyr-tRNA

A U A

Complementary
tRNA anticodon

© 2014 Pearson Education,

 Figure 14.16-
1 Amino acid Tyrosine Tyrosyl-tRNA
and tRNA (Tyr) (amino synthetase
enter active acid)
site.

Tyr-tRNA

A U A

ATP
Complementary
tRNA anticodon AMP  2 P
i

2 Using
ATP,
synthetase
catalyzes
covalent
bonding.
© 2014 Pearson Education,
 Figure 14.16-
1 Amino acid Tyrosine Tyrosyl-tRNA
and tRNA (Tyr) (amino synthetase
enter active acid)
site.

Tyr-tRNA

A U A

ATP
Complementary
tRNA anticodon AMP  2 P
i

2 Using
ATP,
3 Aminoacyl synthetase
tRNA catalyzes
released. covalent
bonding.
© 2014 Pearson Education,
Ribosomes

 Ribosomes facilitate specific coupling of tRNA

anticodons with mRNA codons during protein
synthesis
 The large and small ribosomal are made of
proteins and ribosomal RNAs (rRNAs)
 In bacterial and eukaryotic ribosomes the large and
small subunits join to form a ribosome only when
attached to an mRNA molecule

© 2014 Pearson Education,

Figure 14.17
Growing polypeptide Exit tunnel
tRNA
molecules

Large
subunit
E P
A

Small
subunit

5
mRNA 3

(a) Computer model of functioning ribosome

P site Growing polypeptide
(Peptidyl-tRNA Exit tunnel Amino end Next amino acid
binding site) to be added
to polypeptide
A site (Aminoacyl- chain
E site tRNA binding site)
(Exit site)
E tRNA
E P A Large
mRNA 3
subunit
mRNA
binding site Small 5 Codons
subunit
(b) Schematic model showing binding sites (c) Schematic model with mRNA and tRNA
© 2014 Pearson Education,
 Figure

tRNA Growing polypeptide Exit tunnel

molecules

Large
subunit
EPA

Small
subunit

5
mRNA 3

(a) Computer model of functioning ribosome

© 2014 Pearson Education,

 Figure

P site
(Peptidyl-tRNA Exit tunnel
binding site)

A site (Aminoacyl-
E site tRNA binding site)
(Exit site)
E P A Large
subunit
mRNA
binding site Small
subunit
(b) Schematic model showing binding sites

© 2014 Pearson Education,

 Figure

Growing polypeptide
Amino end Next amino acid
to be added to
polypeptide
chain

E
tRNA
mRNA
3


5 Codons

(c) Schematic model with mRNA and tRNA

© 2014 Pearson Education,
 A ribosome has three binding sites for tRNA
 The P site holds the tRNA that carries the growing
polypeptide chain
 The A site holds the tRNA that carries the next
amino acid to be added to the chain
 The E site is the exit site, where discharged tRNAs
leave the ribosome

© 2014 Pearson Education,

Building a Polypeptide

 The three stages of translation

 Initiation
 Elongation
 Termination
 All three stages require protein “factors” that aid
in the translation process

© 2014 Pearson Education,

Ribosome Association and Initiation of Translation

 The initiation stage of translation brings together

mRNA, a tRNA with the first amino acid, and the
two ribosomal subunits
 A small ribosomal subunit binds with mRNA and a
special initiator tRNA
 Then the small subunit moves along the mRNA until
it reaches the start codon (AUG)

Animation: Translation Introduction

© 2014 Pearson Education,

Figure 14.18

Large
ribosomal
subunit
3 U A C 5 P site
5 A U G
3 Pi
Initiator 
tRNA GTP GDP
E A
mRNA
5 5
Start codon 3 3
Small
mRNA binding site ribosomal Translation initiation complex
subunit
1 Small ribosomal subunit binds 2 Large ribosomal subunit
to mRNA. completes the initiation complex.

© 2014 Pearson Education,

 The start codon is important because it establishes
the reading frame for the mRNA
 The addition of the large ribosomal subunit is last
and completes the formation of the translation
initiation complex
 Proteins called initiation factors bring all these
components together

© 2014 Pearson Education,

Elongation of the Polypeptide Chain

 During elongation, amino acids are added one by

one to the previous amino acid at the C-terminus of
the growing chain
 Each addition involves proteins called elongation
factors and occurs in three steps: codon
recognition, peptide bond formation, and
translocation
 Translation proceeds along the mRNA in a 5 to
3 direction

© 2014 Pearson Education,

 Figure 14.19-
Amino end
of polypeptide
1 Codon recognition

E 3
mRNA
P A
site site GTP
5
GDP  P i

P A

© 2014 Pearson Education,

 Figure 14.19-
Amino end
of polypeptide
1 Codon recognition

E 3
mRNA
P A
site site GTP
5
GDP  P i

P A

2 Peptide bond
formation

P A
© 2014 Pearson Education,
Figure 14.19-3
Amino end
of polypeptide
1 Codon recognition

E 3
Ribosome ready for mRNA P A
next aminoacyl tRNA
site site GTP
5
GDP  P i

E
E
P A
P A
GDP  P i
2 Peptide bond
3 Translocation formation
GTP

© 2014 Pearson Education,

 P A

© 2014 Pearson Education,

Termination of Translation

 Termination occurs when a stop codon in the mRNA

reaches the A site of the ribosome
 The A site accepts a protein called a release factor
 The release factor causes the addition of a water
molecule instead of an amino acid
 This reaction releases the polypeptide, and the
translation assembly then comes apart

© 2014 Pearson Education,

 Figure 14.20-

Release factor

3
5

Stop codon
(UAG, UAA, or UGA)
1 Ribosome reaches a
stop codon on mRNA.

© 2014 Pearson Education,

 Figure 14.20-

Release Free
factor polypeptide

3 3
5 5
Stop codon
(UAG, UAA, or UGA)
1 Ribosome reaches a 2 Release factor
stop codon on mRNA. promotes
hydrolysis.

© 2014 Pearson Education,

 Figure 14.20-

Release factor Free polypeptide

5
3 3
GTP 3
5 5 2

Stop codon 2 GDP P

i
(UAG, UAA, or UGA)
1 Ribosome reaches a 2 Release factor 3 Ribosomal
stop codon on mRNA. promotes subunits and other
hydrolysis. components
dissociate.

© 2014 Pearson Education,

Completing and Targeting the Functional Protein

 Often translation is not sufficient to make a

functional protein
 Polypeptide chains are modified after translation or
targeted to specific sites in the cell

© 2014 Pearson Education,

Protein Folding and Post-Translational
Modifications
 During synthesis, a polypeptide chain spontaneously
coils and folds into its three-dimensional shape
 Proteins may also require post-translational
modifications before doing their jobs

© 2014 Pearson Education,

Targeting Polypeptides to Specific Locations

 Two populations of ribosomes are evident in cells:

free ribosomes (in the cytosol) and bound
ribosomes (attached to the ER)
 Free ribosomes mostly synthesize proteins that
function in the cytosol
 Bound ribosomes make proteins of the
endomembrane system and proteins that are
secreted from the cell

© 2014 Pearson Education,

 Polypeptide synthesis always begins in the cytosol
 Synthesis finishes in the cytosol unless the
polypeptide signals the ribosome to attach to the
ER
 Polypeptides destined for the ER or for secretion
are marked by a signal peptide

© 2014 Pearson Education,

 A signal-recognition particle (SRP) binds to the
signal peptide
 The SRP brings the signal peptide and its ribosome
to the ER

© 2014 Pearson Education,

Figure 14.21

1 2 3 4 5 6
Polypeptide SRP SRP SRP Signal- Completed
synthesis binds to binds to detaches cleaving polypeptide
begins. signal receptor and enzyme folds into
peptide. protein. polypeptide cuts off final
synthesis signal conformation.
resumes. peptide.
Ribosome
mRNA

Signal ER
peptide membrane
SRP Signal
peptide Protein
removed

CYTOSOL SRP receptor

protein
ER LUMEM
Translocation complex
© 2014 Pearson Education,
Making Multiple Polypeptides in Bacteria and
Eukaryotes
 In bacteria and eukaryotes multiple ribosomes
translate an mRNA at the same time
 Once a ribosome is far enough past the start codon,
another ribosome can attach to the mRNA
 Strings of ribosomes called polyribosomes (or
polysomes) can be seen with an electron
microscope

© 2014 Pearson Education,

Figure 14.22
Growing Completed
polypeptides polypeptide

Incoming
ribosomal
subunits

Start of
mRNA End of mRNA
(5 (3 end)
end)
(a) Several ribosomes simultaneously translating one
mRNA molecule

Ribosomes

mRNA

© 2014 Pearson Education,

 (b) A large polyribosome in a
bacterial cell (TEM) 0.1 m

© 2014 Pearson Education,

 Figure

Growing Completed
polypeptides polypeptide

Incoming
ribosomal
subunits

Start of
mRNA End of mRNA
(5 (3 end)
end)
(a) Several ribosomes simultaneously translating
one mRNA molecule

© 2014 Pearson Education,

 Figure

Ribosomes

mRNA

(b) A large polyribosome in a 0.1 m

bacterial cell (TEM)

© 2014 Pearson Education,

 Bacteria and eukaryotes can also transcribe
multiple mRNAs form the same gene
 In bacteria, the transcription and translation can
take place simultaneously
 In eukaryotes, the nuclear envelope separates
transcription and translation

© 2014 Pearson Education,

 Figure
RNA polymerase

DNA
mRNA
Polyribosome

Direction of 0.25 m
RNA transcription
polymerase
DNA

Polyribosome
Polypeptide
(amino end)

Ribosome

mRNA (5 end)

© 2014 Pearson Education,
 Figure

RNA polymerase

DNA
mRNA

Polyribosome

0.25 m

© 2014 Pearson Education,

 Figure
TRANSCRIPTION DNA

3

RNA
5 RNA
polymerase
transcript Exon
RNA
PROCESSING RNA
transcript
(pre-mRNA)
Intron Aminoacyl-tRNA
synthetase
NUCLEUS
Amino
acid
AMINO ACID
CYTOPLASM tRNA ACTIVATION

mRNA

A 3
P Aminoacyl
E Ribosomal (charged)
subunits tRNA

TRANSLATION
E A
A A A Anticodon
U G G U U U A U G

Codon

© 2014 Pearson Education,

 Figure Ribosome

© 2014 Pearson Education,

 Figure

DNA
TRANSCRIPTION

3

5 RNA RNA
transcript polymerase
RNA Exon
PROCESSING
RNA
transcript
(pre-mRNA)
Intron Aminoacyl-tRNA
synthetase
NUCLEUS
Amino
acid
AMINO ACID
CYTOPLASM tRNA ACTIVATION

mRNA
Aminoacyl
(charged)
© 2014 Pearson Education,
 Figure
t
R
N
A

© 2014 Pearson Education,

 Figure

mRNA Growing polypeptide

3
A
P Aminoacyl
E Ribosomal (charged)
subunits tRNA

TRANSLATION
E A
A A A Anticodon
U G G U U U A U G

Codon
Ribosome

© 2014 Pearson Education,

Concept 14.5: Mutations of one or a few
nucleotides can affect protein structure and
function
 Mutations are changes in the genetic material of a
cell or virus
 Point mutations are chemical changes in just one
or a few nucleotide pairs of a gene
 The change of a single nucleotide in a DNA
template strand can lead to the production of an
abnormal protein

© 2014 Pearson Education,

Figure 14.25

Wild-type hemoglobin Sickle-cell hemoglobin

Wild-type hemoglobin DNA Mutant hemoglobin DNA
3 C T C 5 3 C A C 5
5 G A G 3 5 G T G 3

mRNA mRNA
5 G A G 3 G U G 3

5

Normal hemoglobin Sickle-cell hemoglobin

Glu Val
© 2014 Pearson Education,
Types of Small-Scale Mutations

 Point mutations within a gene can be divided into

two general categories
 Nucleotide-pair substitutions
 One or more nucleotide-pair insertions or deletions

© 2014 Pearson Education,

Figure 14.26
Wild type
DNA template
3 T A C T T C A A C C G A T T 5
strand A
5 A T G A A G T T T G G C T A A 3
mRNA 5 A G A A G U U U G G C U A A 3
U
Protein Met Lys Phe Gly
Stop
Amino end Carboxyl end
(a) Nucleotide-pair substitution (b) Nucleotide-pair insertion or deletion
A instead of Extra A
G
3 T A
C T T C A A A C C A A T T 3 T A C A T T C A A A C C G A T T 5
5
5 A T G A A G T T T G G T T A A 5 A G T A A G T T T G G C T A A 3
3 T
U instead of C Extra U
5 A U G A A G U U U G G U U A A 5 A G U A A G U U U G G U U A A 3
3 U
Met Lys Phe Gly Met
Stop Stop
Silent (no effect on amino acid sequence) Frameshift causing immediate nonsense
(1 nucleotide-pair insertion)
T instead of C A missing
3 T
A C T T C A A A T C G A T T 3 T C T T C A A C C G A T T 5
5 A
5 A T G A A G T T T A G C T A A 5 A G A A G T T G G C T A A 3
3 T
A instead of G U missing
5 A U G A A G U U U A G C U A A 5 A G A A G U U G G G U A A 3
3 U
Met Lys Phe Ser Met Lys Leu Ala
Stop
Missense Frameshift causing extensive missense
(1 nucleotide-pair deletion)
© 2014 Pearson Education,
 A instead of T T T C missing
3 T
A C A T C A A A C C G A T T 3 T A A A A C C G A T T 5
5 C
5 A T G T A G T T T G G C T A A 5 A T T T T G G C T A A 3
3 G
U instead of A A A G missing
5 A U G U A G U U U G G U U A A 5 A U U U U G G C U A A 3
3 G
Met Met Phe Gly
Stop Stop
Nonsense No frameshift, but one amino acid missing
(3 nucleotide-pair deletion)

© 2014 Pearson Education,

Substitutions

 A nucleotide-pair substitution replaces one

nucleotide and its partner with another pair of
nucleotides
 Silent mutations have no effect on the amino acid
produced by a codon because of redundancy in the
genetic code

© 2014 Pearson Education,

Figure 14.26a

Wild type
DNA template 3 T C T T C A A A C C G A T T 5
strand
A
5 T G A A G T T T G G C T A 3
mRNA A A
U G A A G U U A 3
5 U G G C U
A A
Protein Met Lys Phe Gly
Stop
Amino end
Carboxyl
end

Nucleotide-pair substitution: silent

A instead of G
3 T A
C T T C A A A C C A A T T 5
5 A T G A A G T T G G T T A A 3
T
© 2014 Pearson Education,
 U instead of C
5 A U G A G U U U G G U U A 3
A A
Met Lys Phe Gly
Stop

© 2014 Pearson Education,

 Missense mutations still code for an amino acid,
but not the correct amino acid
 Substitution mutations are usually missense
mutations
 Nonsense mutations change an amino acid codon
into a stop codon, nearly always leading to a
nonfunctional protein

Animation: Protein Synthesis

© 2014 Pearson Education,

 Figure

Wild type
DNA template 3 T A C T T C A A A C C G A T T 5
strand
5 T G A A G T T T G G C T A 3
mRNA A A
U G A A G U U A 3
5 U G G C U
A A
Protein Met Lys Phe Gly
Stop
Amino end
Carboxyl
end

Nucleotide-pair substitution: missense

T instead of C
3 T A C T T C A A A T C G A T T 5
5 A T G A A G T T T A G C T A A
3
A instead of G
5 A U G A A
G U U U A G C
© 2014 Pearson Education,
 Figure
U A A 3
Met Lys Phe Ser
Stop

© 2014 Pearson Education,

 Figure

Wild type
DNA template 3 T A C T T C A A A C C G A T T 5
strand 5 A T G A A G T T T G G C T A A 3
mRNA 5 U G A A G U U G G C U A 3
A U A
Protein Met Lys Phe Gly
Stop
Amino end
Carboxyl end

Nucleotide-pair substitution: nonsense

A instead of T
3 T A C A T C A A A C C G A T T
5
5 A T G T A G T T G G C A A 3
T T
U instead of A
5 A U G U G U U U G G C U A A 3
A

© 2014 Pearson Education,

 Figure
Met
Stop

© 2014 Pearson Education,

Insertions and Deletions

 Insertions and deletions are additions or losses of

nucleotide pairs in a gene
 These mutations have a disastrous effect on the
resulting protein more often than substitutions do
 Insertion or deletion of nucleotides may alter the
reading frame of the genetic message, producing a
frameshift mutation

© 2014 Pearson Education,

 Figure

Wild type
DNA template 3 T A C T T C A A A C C G A T T 5
strand 5 A T G A A G T T T G G C T A A 3
mRNA 5 U G A A G U U U G G C U A 3
A A
Protein Met Lys Phe Gly
Stop
Amino end
Carboxyl
end

Nucleotide-pair insertion: frameshift causing immediate nonsense

Extra A
3 T A C A T T C A A A C C G A T T 5
5 A T G T A A G T T T G G C T A A 3
Extra U
5 A U G U A A G U U U G G C U A A 3
Met
Stop
© 2014 Pearson Education,
 Figure

Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3
mRNA 5 A U G A A G U U U G G C U A A 3
Protein Met Lys Phe Gly
Stop
Amino end
Carboxyl end

Nucleotide-pair deletion: frameshift causing extensive missense

A missing
3 T A C T T C A A C C G A T T 5
5 A T G A A T T G G C T A A 3
G
U missing
5 A U G A A G U U G G C U A A 3
Met Lys Leu Ala

© 2014 Pearson Education,

 Figure

Wild type
DNA template 3 T C T T C A A A C C G A T T 5
strand
A
5 T G A A G T T T G G C T A 3
mRNA A A
U G A A G U U A 3
5 U G G C U
A A
Protein Met Lys Phe Gly
Stop
Amino end
Carboxyl
end

3 nucleotide-pair deletion: no frameshift, but one amino acid

missing
T T C
missing
3 T A C A A A C C A T T 5
G
5 A T G T T

© 2014 Pearson Education,

 Figure
T G G C T A A 3
A A G
missing
5 A U G U U U G G C U A A 3
Met Phe Gly
Stop

© 2014 Pearson Education,

Mutagens

 Spontaneous mutations can occur during DNA

replication, recombination, or repair
 Mutagens are physical or chemical agents that can
cause mutations
 Researchers have developed methods to test the
mutagenic activity of chemicals
 Most cancer-causing chemicals (carcinogens) are
mutagenic, and the converse is also true

© 2014 Pearson Education,

What Is a Gene? Revisiting the Question

 The definition of a gene has evolved through the

history of genetics
 We have considered a gene as
 A discrete unit of inheritance
 A region of specific nucleotide sequence in a
chromosome
 A DNA sequence that codes for a specific polypeptide
chain

© 2014 Pearson Education,

 A gene can be defined as a region of DNA that can
be expressed to produce a final functional product,
either a polypeptide or an RNA molecule

© 2014 Pearson Education,

Figure 14.UN05a

thrA
lacA
lacY
lacZ
lacl
recA
galR
met J
lexA
trpR
5 3

−1

−2
−1

−1

−1
−1

−1

−1

−1

−1

−7
−6
−5
−4
5

−9
−8

−3
8

−1
1

© 2014 Pearson Education,

 ©

Figure
2014 Pearson Education,

5
−18
−17
−16
−15
−14
−13
−12
−11
−10
−9
−8
−7
−6
−5
−4
−3
−2
−1
0
1
2
3
4
5
6
7
8
3
 ©

Figure
−18
2014 Pearson Education,

−17
−16
−15
−14
−13
−12
−11
−10
−9
−8
−7
−6
−5
−4
−3
−2
−1
0
1
2
3
4
5
6
7
8
 Figure

Transcription unit

Promoter

5 3
3
3 5
5
Template strand
RNA polymerase of DNA
RNA transcript

© 2014 Pearson Education,

 Figure

Pre-mRNA
5 Cap
Poly-A tail

mRNA

© 2014 Pearson Education,

 Figure

Polypeptide

tRNA Amino
acid

E A Anti-
codon
Codon
Ribosome mRNA

© 2014 Pearson Education,

 Figure

© 2014 Pearson Education,