Biology

Sylvia S. Mader
Michael Windelspecht

Chapter 12
Molecular Biology
of the Gene
Lecture Outline

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1
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 Outline
• 12.1 The Genetic Material
• 12.2 Replication of DNA
• 12.3 The Genetic Code of Life
• 12.4 First Step: Transcription
• 12.5 Second Step: Translocation
• 12.6 Structure of the Eukaryotic Chromosome

2
 12.1 The Genetic Material
• Frederick Griffith investigated virulence of
Streptococcus pneumoniae
 Concluded that virulence could be passed
from a dead strain to a nonvirulent living strain
 Transformation
• Further research by Avery et al.
 Discovered that DNA is the transforming
substance
 DNA from dead cells was being incorporated
into the genome of living cells
3
 The Genetic Material
• Griffith’s Transformation Experiment
 Mice were injected with two strains of
pneumococcus: an encapsulated (S) strain
and a non-encapsulated (R) strain.
• The S strain is virulent (the mice died); it has a
mucous capsule and forms “shiny” colonies.
• The R strain is not virulent (the mice lived); it has
no capsule and forms “dull” colonies.

4
 Griffith’s Transformation
Experiment
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capsule

Injected live
Injected live R strain has Injected heat- Injected heat-killed
S strain has no capsule killed S strain S strain plus live
capsule and and mice does not cause R strain causes
causes mice do not die. mice to die. mice to die. Live S strain is
to die. withdrawn from
dead mice.

a. b. c. d.

5
 The Genetic Material
• Transformation of organisms today:
 Result is the so-called genetically modified
organisms (GMOs)
• Invaluable tool in modern biotechnology today
• Commercial products that are currently much used
• Green fluorescent protein (GFP) can be used as a
marker
– A jellyfish gene codes for GFP
– The jellyfish gene is isolated and then transferred to a
bacterium, or the embryo of a plant, pig, or mouse.
– When this gene is transferred to another organism, the
organism glows in the dark
6
Transformation of Organisms

7
 The Genetic Material
• DNA contains:
 Two Nucleotides with purine bases
• Adenine (A)
• Guanine (G)
 Two Nucleotides with pyrimidine bases
• Thymine (T)
• Cytosine (C)

8
 The Genetic Material
• Chargaff’s Rules:
 The amounts of A, T, G, and C in DNA:
• Are constant among members of the same species
• Vary from species to species
 In each species, there are equal amounts of:
• A and T
• G and C
 All this suggests that DNA uses
complementary base pairing to store genetic
information
 Each human chromosome contains, on
average, about 140 million base pairs
 The number of possible nucleotide sequences
is 4140,000,000
9
Nucleotide Composition of DNA Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
O
NH2
C CH3
adenine C thymine HN C
C N
(A) N (T)
CH nitrogen-containing O C CH
HC C base
N N
O N
O
5 5
HO P O CH2 O HO P O CH2 O
O C H H C1 O C H H C1
4 4 sugar = deoxyribose
H C C H H C C H NH2
3 2 O 3 2
OH H OH H C
guanine C cytosine N CH
C N
(G) HN (C)
CH C CH
O
H2N C C N N
O N
O
5 5
phosphate HO P O CH2 O HO P O CH2 O
O C H H C1 O C H H C1
4 4
H C C H H C
2 C H 2
3 3
a. Purine nucleotides OH H b. Pyrimidine nucleotides OH H

DNA Composition in Various Species (%)

Species A T G C

Homo sapiens (human) 31.0 31.5 19.1 18.4

Drosophila melanogaster (fruit fly) 27.3 27.6 22.5 22.5
Zea mays (corn) 25.6 25.3 24.5 24.6
Neurospora crassa (fungus) 23.0 23.3 27.1 26.6
Escherichia coli (bacterium) 24.6 24.3 25.5 25.6
10
Bacillus subtilis (bacterium) 28.4 29.0 21.0 21.6
c. Chargaff’s data
 The Genetic Material
• X-Ray diffraction:
 Rosalind Franklin studied the structure of DNA using
X-rays.
 She found that if a concentrated, viscous solution of
DNA is made, it can be separated into fibers.
 Under the right conditions, the fibers can produce an
X-ray diffraction pattern
• She produced X-ray diffraction photographs.
• This provided evidence that DNA had the following features:
– DNA is a helix.
– Some portion of the helix is repeated.

11
 X-Ray Diffraction of DNA
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Rosalind Franklin

diffraction pattern
diffracted
X-rays
a.
X-ray beam

Crystalline
DNA

c.
b.
© Photo Researchers, Inc.; c: © Science Source/Photo Researchers, Inc.

12
 The Genetic Material
• The Watson and Crick Model (1953)
 Double helix model is similar to a twisted ladder
• Sugar-phosphate backbones make up the sides

• Hydrogen-bonded bases make up the rungs

 Complementary base pairing ensures that a purine is

always bonded to a pyrimidine (A with T, G with C)

 Received a Nobel Prize in 1962

13
Watson and Crick Model of DNACopyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
3.4 nm
0.34 nm

2 nm

b.

d.

C
a. sugar-phosphate
G
T backbone
5′ end 3′ end
P
A P
G C
S S A
T
P
P P
T A S G
S P C
3′ end 5′ end P
complementary C
c.
base pairing

G
P

hydrogen
sugar bonds 14
a: © Photodisk Red/Getty RF; d: © A. Barrington Brown/Photo Researchers
 12.2 Replication of DNA
• DNA replication is the process of copying
a DNA molecule.
• Semiconservative replication - each
strand of the original double helix (parental
molecule) serves as a template (mold or
model) for a new strand in a daughter
molecule.

15
Semiconservative Replication
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
5′ 3′
G
C G
C G
A T
A

region of parental T A
DNA double helix
C G
DNA
A T polymerase
A enzyme
G
C G
C G
A

region of
replication:
new nucleotides
are pairing
with those of
parental strands

region of
completed
replication
3′
new old
strand strand

5′ daughter DNA double helix

old new 16
strand strand
daughter DNA double helix
 Replication of DNA
• Replication requires the following steps:
 Unwinding, or separation of the two strands of the
parental DNA molecule

 Complementary base pairing between a new

nucleotide and a nucleotide on the template strand

 Joining of nucleotides to form the new strand

• Each daughter DNA molecule contains one old strand and one
new strand

17
 Replication of DNA
• Prokaryotic Replication
 Bacteria have a single circular loop of DNA
 Replication moves around the circular DNA molecule
in both directions
 Produces two identical circles
 The process begins at the origin of replication
 Replication takes about 40 minutes, but the cell
divides every 20 minutes
• A new round of replication can begin before the previous
round is completed

18
 Replication of DNA
• Eukaryotic Replication
 DNA replication begins at numerous points
along each linear chromosome
 DNA unwinds and unzips into two strands
 Each old strand of DNA serves as a template
for a new strand
 Complementary base-pairing forms a new
strand paired with each old strand
• Requires enzyme DNA polymerase

19
 Replication of DNA
• Eukaryotic Replication
 Replication bubbles spread bidirectionally until
they meet
 The complementary nucleotides are joined to form
new strands. Each daughter DNA molecule
contains an old strand and a new strand.
 Replication is semiconservative:
• One original strand is conserved in each daughter
molecule, i.e., each daughter double helix has one
parental strand and one new strand.

20
Prokaryotic versus Eukaryotic Replication
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origin

replication is
complete

replication is
occurring in
two directions

a. Replication in prokaryotes
replication fork replication bubble

parental strand

new DNA
duplexes
21
daughter strand
b. Replication in eukaryotes
Aspects of DNA Replication Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

OH P Is attached here
base is attached here
5′
CH2
O OH
4′ C C
H H 1′ 1
H H
C
3′ C 2′
OH H
De ox yrib os e m o lec u le

2 DNA polymerase
5′ end attaches a new
nucleotide to the
P 3 ′ carbon of the
previous nucleotide.

P
P

G C 3′ e nd
P

P
P C G

5′
P 3′
P T A

C P template
P G
strand

P
3 ′end 5′ end 4 leading
DNA polymerase
template strand new strand
new strand
Direction of replication 3′ 3 helicase at replication fork
RNA primer
template 6 Okazaki fragment
lagging strand
5
strand 3′

5′
5′ parental DNA helix
22
7 DNA ligase DNA polymerase
3′
Replication fork introduces complications
 Replication of DNA
• Accuracy of Replication
 DNA polymerase is very accurate, yet makes
a mistake about once per 100,000 base pairs.
• Capable of identifying and correcting errors

23
12.3 The Genetic Code of Life
• Genes Specify Enzymes
 Beadle and Tatum:
• Experiments on the fungus Neurospora
crassa
• Proposed that each gene specifies the
synthesis of one enzyme
• One-gene-one-enzyme hypothesis

24
 The Genetic Code of Life
• The mechanism of gene expression
 DNA in genes specify information, but
information is not structure and function
 Genetic information is expressed into
structure and function through protein
synthesis
• The expression of genetic information into
structure and function:
 DNA in a gene determines the sequence of
nucleotides in an RNA molecule
 RNA controls the primary structure of a
protein 25
The Central Dogma of Molecular Biology

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

3'
5'

A G C G A C C C C

DNA

T C G C T G G G G

3' 5'

26
The Central Dogma of Molecular Biology
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nontemplate strand
5 3
A G C G A C C C C

DNA T C G C T G G G G
3 5
template strand
transcription
in nucleus

5 3

mRNA A G C G A C C C C

27
The Central Dogma of Molecular Biology
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nontemplate strand
5 3

A G C G A C C C C

DNA T C G C T G G G G
3 5
template strand
transcription
at ribosome

5 3
A G C G A C C C C
mRNA

codon 1 codon 2 codon 3 28

The Central Dogma of Molecular Biology
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
nontemplate strand
5 3

A G C G A C C C C

DNA T C G C T G G G G

3 5
template strand
transcription
in nucleus

5 3

A G C G A C C C C
mRN A

translation codon 1 codon 2 codon 3

at ribosome

O O O

polypeptide N C C N C N C C

R1 R2 R3
29
Serine Aspartate Proline
 The Genetic Code of Life
• RNA is a polymer of RNA nucleotides
 RNA nucleotides contain the sugar ribose instead of
deoxyribose
 RNA nucleotides are of four types: uracil (U),
adenine (A), cytosine (C), and guanine(G)
 Uracil (U) replaces thymine (T) of DNA
• Types of RNA
• Messenger (mRNA) - Takes a message from DNA in the
nucleus to ribosomes in the cytoplasm
• Ribosomal (rRNA) - Makes up ribosomes, which read the
message in mRNA
• Transfer (tRNA) - Transfers the appropriate amino acid to
the ribosomes for protein synthesis

30
 Structure of RNA
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5′ end

P
G

U S

A P
base is
uracil instead
C of thymine
S

ribose one nucleotide

3′ end
31
RNA Structure Compared to
DNA structure

32
 The Genetic Code of Life
• The unit of the genetic code consists of codons,
each of which is a unique arrangement of symbols
• Each of the 20 amino acids found in proteins is
uniquely specified by one or more codons
 The symbols used by the genetic code are the mRNA bases
• Function as “letters” of the genetic alphabet
• Genetic alphabet has only four “letters” (U, A, C, G)
 Codons in the genetic code are all three bases (symbols)
long
• Function as “words” of genetic information
• Permutations:
– There are 64 possible arrangements of four symbols taken
three at a time
– Often referred to as triplets
• Genetic language only has 64 “words”

33
Messenger RNA Codons
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

First Second Base Third

Base U C A G Base
UUU UCU UAU UGU
U
phenylalanine serine tyrosine cysteine
UUC UCC UAC UGU
C
phenylalanine serine tyrosine cysteine
U
UUA UCA UAA UGA
A
leucine serine stop stop
UUG UCG UAG UGG
G
leucine serine stop tryptophan
CUU CCU CAU CGU
U
leucine proline histidine arginine
CUC CCC CAC CGC
C
leucine proline histidine arginine
C
CUA CCA CAA CGA
A
leucine proline glutamine arginine
CUG CCG CAG CGG G
leucine proline glutamine arginine
AUU ACU AAU AGU U
isoleucine threonine asparagine serine
AUC ACC AAC AGC C
isoleucine threonine asparagine serine
A
AUA ACA AAA AGA
A
isoleucine threonine lysine arginine
AUG (start) ACG AAG AGG G
methionine threonine lysine arginine
GUU GCU GAU GGU U
valine alanine aspartate glycine
GUC GCC GAC GGC C
valine alanine aspartate glycine
G
GUA GCA GAA GGA A
valine alanine glutamate glycine
34
GUG GCG GAG GGG G
valine alanine glutamate glycine
 The Genetic Code of Life
• Properties of the genetic code:
 Universal
• With few exceptions, all organisms use the code the same
way
• Encode the same 20 amino acids with the same 64 triplets
 Degenerate (redundant)
• There are 64 codons available for 20 amino acids
• Most amino acids encoded by two or more codons
 Unambiguous (codons are exclusive)
• None of the codons code for two or more amino acids
• Each codon specifies only one of the 20 amino acids
 Contains start and stop signals
• Punctuation codons
• Like the capital letter we use to signify the beginning of a
sentence, and the period to signify the end
35
 12.4 First Step: Transcription
• Transcription
 A segment of DNA serves as a template for
the production of an RNA molecule
 The gene unzips and exposes unpaired bases
 Serves as template for mRNA formation
 Loose RNA nucleotides bind to exposed DNA
bases using the C=G and A=U rule
 When entire gene is transcribed into mRNA,
the result is a pre-mRNA transcript of the
gene
 The base sequence in the pre-mRNA is
complementary to the base sequence in DNA
36
 First Step: Transcription
• A single chromosome consists of one very long molecule encoding
hundreds or thousands of genes
• The genetic information in a gene describes the amino acid
sequence of a protein
 The information is in the base sequence of one side (the “sense” strand)
of the DNA molecule
 The gene is the functional equivalent of a “sentence”
• The segment of DNA corresponding to a gene is unzipped to expose
the bases of the sense strand
 The genetic information in the gene is transcribed (rewritten) into an
mRNA molecule
 The exposed bases in the DNA determine the sequence in which the
RNA bases will be connected together
 RNA polymerase connects the loose RNA nucleotides together
• The completed transcript contains the information from the gene, but
in a mirror image, or complementary form

37
 Transcription
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3′ 5′

terminator

gene strand template

strand

3′ C
direction of
polymerase
movement
T
RNA
polymerase

DNA
template
strand

mRNA
transcript

promoter

5′

38
5′ 3′ to RNA processing
RNA Polymerase
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a. 200m

spliceosome

DNA

RNA
polymerase

RNA
transcripts

b.
© Oscar L. Miller/Photo Researchers, Inc.
39
 First Step: Transcription
• Pre-mRNA is modified before leaving the
eukaryotic nucleus.
 Modifications to the ends of the primary
transcript:
• Cap on the 5′ end
– The cap is a modified guanine (G) nucleotide
– Helps a ribosome determine where to attach when
translation begins
• Poly-A tail of 150-200 adenines on the 3′ end
– Facilitates the transport of mRNA out of the nucleus
– Inhibits degradation of mRNA by hydrolytic enzymes.
40
 First Step: Transcription
• Pre-mRNA, is composed of exons and introns.
 The exons will be expressed,
 The introns, occur in between the exons.
• Allows a cell to pick and choose which exons will go into a
particular mRNA
 RNA splicing:
• Primary transcript consists of:
– Some segments that will not be expressed (introns)
– Segments that will be expressed (exons)
• Performed by spliceosome complexes in nucleoplasm
– Introns are excised
– Remaining exons are spliced back together

• Result is a mature mRNA transcript

41
 First Step: Transcription
• In prokaryotes, introns are removed by
“self-splicing”—that is, the intron itself has
the capability of enzymatically splicing
itself out of a pre-mRNA

42
Messenger RNA Processing in Eukaryotes

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DNA

43
Messenger RNA Processing in Eukaryotes

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exon exon exon

DNA
intron intron

44
Messenger RNA Processing in Eukaryotes
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exon exon exon

DNA
intron intron
transcription

exon exon exon

pre-mRNA
5 intron intron 3

45
Messenger RNA Processing in Eukaryotes
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exon exon exon

DNA
intron intron
transcription

exon exon exon

pre-mRNA
5 intron intron 3

exon exon exon

5 3
cap intron intron poly-A tail 46
Messenger RNA Processing in Eukaryotes
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exon exon exon

DNA
intron intron
transcription

exon exon exon

pre-mRNA
5 intron intron 3

exon exon exon

5 3
cap intron intron poly-A tail

spliceosome

exon exon exon

5 3 47
cap poly-A tail
Messenger RNA Processing in Eukaryotes
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exon exon exon

DNA
intron intron
transcription

exon exon exon

pre-mRNA
5 intron intron 3

exon exon exon

5 3
cap intron intron poly-A tail

spliceosome

exon exon exon

5 3
cap poly-A tail
pre-mRNA
splicing

intron RNA

mRNA
5 3 48
cap poly-A tail
Messenger RNA Processing in Eukaryotes
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exon exon exon

DNA
intron intron
transcription

exon exon exon

pre-mRNA
5 intron intron 3

exon exon exon

5 3
cap intron intron poly-A tail

spliceosome

exon exon exon

5 3
cap poly-A tail
pre-mRNA
splicing

intron RNA

mRNA
5 3
cap poly-A tail

49
cytoplasm
Messenger RNA Processing in Eukaryotes
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exon exon exon

DNA
intron intron
transcription

exon exon exon

pre-mRNA
5 intron intron 3

exon exon exon

5 3
cap intron intron poly-A tail

spliceosome

exon exon exon

5 3
cap poly-A tail
pre-mRNA
splicing

intron RNA
mRNA
5 3
cap poly-A tail
nuclear pore
in nuclear envelope
nucleus
50
cytoplasm
 First Step: Transcription
• Functions of introns:
 As organismal complexity increases;
• The number of protein-coding genes does not keep pace
• But the proportion of the genome that is introns increases
 Possible functions of introns:
• Exons might combine in various combinations
– Would allow different mRNAs to result from one segment of DNA
• Introns might regulate gene expression
• Introns may encourage crossing-over during meiosis
 Exciting new picture of the genome is
emerging

51
12.5 Second Step: Translation
• Translation
 The sequence of codons in the mRNA at a ribosome directs the sequence of
amino acids into a polypeptide
 A nucleic acid sequence is translated into a protein sequence
• tRNA molecules have two binding sites:
 One associates with the mRNA transcript
 The other associates with a specific amino acid
 Each of the 20 amino acids in proteins associates with one or more of 64
types of tRNA
• Translation
 An mRNA transcript associates with the rRNA of a ribosome in the
cytoplasm or a ribosome associated with the rough endoplasmic reticulum
 The ribosome “reads” the information in the transcript
 Ribosome directs various types of tRNA to bring in their specific amino acid
“fares”
 The tRNA specified is determined by the code being translated in the mRNA
transcript

52
 Second Step: Translation
• tRNA molecules come in 64 different kinds
• All are very similar except that
 One end bears a specific triplet (of the 64
possible) called the anticodon
 The other end binds with a specific amino acid
type
 tRNA synthetases attach the correct amino acid
to the correct tRNA molecule
• All tRNA molecules with a specific anticodon
will always bind with the same amino acid
53
Structure of a tRNA Molecule
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amino
acid
leucine

3’
5’

hydrogen
bonding

G A A

54
Structure of a tRNA Molecule
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amino
acid
leucine

3’
5’

hydrogen
bonding

amino acid end

G A A

55
Structure of a tRNA Molecule
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amino
acid
leucine

3’
5’

hydrogen
bonding

amino acid end

anticodon

G A A anticodon end

56
 Structure of a tRNA Molecule Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

amino
acid
leucine

3’
5’

hydrogen
bonding

amino acid end

anticodon

G A A anticodon end

C A G U C C U U C C U C
mRNA
5’ 3’

57
 Structure of a tRNA Molecule Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

amino
acid
leucine

3’
5’

hydrogen
bonding

amino acid end

anticodon

G A A anticodon end

C A G U C C U U C C U C
mRNA
5’ 3’
codon
a. b. 58
 Second Step: Translation
• Ribosomes
 Ribosomal RNA (rRNA):
• Produced from a DNA template in the nucleolus of a nucleus
• Combined with proteins into large and small ribosomal
subunits

 A completed ribosome has three binding sites to

facilitate pairing between tRNA and mRNA
• The E (for exit) site
• The P (for peptide) site, and
• The A (for amino acid) site

59
 Ribosome Structure and Function
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large subunit

5 3

mRNA
tRNA binding
sites

small subunit
a. Structure of a ribosome b. Binding sites of ribosome outgoing
tRNA

polypeptide
incoming
tRNA

mRNA

c. Function of ribosomes d. Polyribosome

Courtesy Alexander Rich 60

 Second Step: Translation
• Initiation:
 Components necessary for initiation are:
• Small ribosomal subunit
• mRNA transcript
• Initiator tRNA, and
• Large ribosomal subunit
• Initiation factors (special proteins that bring the
above together)
 Initiator tRNA:
• Always has the UAC anticodon
• Always carries the amino acid methionine
• Capable of binding to the P site

61
 Second Step: Translation
• Small ribosomal subunit attaches to
mRNA transcript

 Beginning of transcript always has the START

codon (AUG)

• Initiator tRNA (UAC) attaches to P site

• Large ribosomal subunit joins the small

subunit
62
 Initiation
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amino acid methionine

Met

initiator tRNA

5 mRNA E site P site A site

3 Met

small ribosomal subunit

large ribosomal subunit U AC

AUG

5 start codon 3

A small ribosomal subunit

binds to mRNA; an initiator
tRNA pairs with the mRNA
start codon AUG. The large ribosomal subunit
completes the ribosome.
Initiator tRNA occupies the
P site. The A site is ready
for the next tRNA.

63
Initiation
 Second Step: Elongation
• “Elongation” refers to the growth in length
of the polypeptide
• RNA molecules bring their amino acid
fares to the ribosome
 Ribosome reads a codon in the mRNA
• Allows only one type of tRNA to bring its amino
acid
• Must have the anticodon complementary to the
mRNA codon being read
• The incoming tRNA joins the ribosome at its A site
 Methionine of the initiator tRNA is connected
to the amino acid of the 2nd tRNA by a peptide64
bond
 Second Step: Elongation
• The second tRNA moves to P site
(translocation)
• The spent initiator tRNA moves to the E site and
exits
• The ribosome reads the next codon in the
mRNA
 Allows only one type of tRNA to bring its amino acid
• Must have the anticodon complementary to the mRNA codon
being read
• Joins the ribosome at its A site
 The dipeptide on the 2nd amino acid is connected to
65
the amino acid of the 3rd tRNA by a peptide bond
 Elongation
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Met
peptide
Ser bond

Ala

Trp

Val

C A U
G U A G A C

5 3
66
 Elongation
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asp
Met
peptide tRNA
Ser bond

Ala C U G

Trp anticodon

Val

C A U
G U A G A C

3
5

1 A tRNA–amino acid
approaches the
ribosome and binds
at the A site. 67
 Elongation
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asp
Met Met
peptide tRNA
Ser bond Ser
Ala C UG Ala
Trp anticodon Trp
Val Val Asp

C AU C A U C UG
G UA GA C G UA GA C

3 3
5 5

1 2
A tRNA–amino acid Two tRNAs can be at a
approaches the ribosome at one time;
ribosome and binds the anticodons are
at the A site. paired to the codons. 68
 Elongation
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Met
asp
Met Met Ser
peptide tRNA
Ser bond Ser Ala
Ala C U G Ala Trp
peptide
Trp anticodon Trp Val bond
Val Val Asp Asp

C A U C A U C U G C A U CU G
G U A G A C GU A GA C G U A GA C
3
5 3 5 5 3

1 A tRNA–amino acid 2 Two tRNAs can be at a 3 Peptide bond formation

approaches the ribosome at one time; attaches the peptide
ribosome and binds the anticodons are chain to the newly
at the A site. paired to the codons. arrived amino acid.

69
 Elongation
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Met Met
asp
Met Met Ser
peptide tRNA Ser
Ser bond Ser Ala
Ala
Ala C U G Ala Trp Trp
peptide
Trp anticodon Trp Val bond Val
Val Val Asp
Asp Asp

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

3
5 3 5 5 3 5 3

1 A tRNA–amino acid 2 Two tRNAs can be at a 3 Peptide bond formation 4 The ribosome moves forward; the
approaches the ribosome at one time; attaches the peptide “empty” tRNA exits from the E site;
ribosome and binds the anticodons are chain to the newly the next amino acid–tRNA complex
at the A site. paired to the codons. arrived amino acid. is approaching the ribosome.

70
 Elongation
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Met Met
asp
Met Met Ser Thr
peptide tRNA Ser
Ser bond Ser Ala
Ala
Ala C U G Ala Trp Trp
peptide
Trp anticodon Trp Val bond Val
Val Val Asp
Asp Asp

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

3
5 3 5 5 3 5 3

1 A tRNA–amino acid 2 Two tRNAs can be at a 3 Peptide bond formation 4 The ribosome moves forward; the
approaches the ribosome at one time; attaches the peptide “empty” tRNA exits from the E site;
ribosome and binds the anticodons are chain to the newly the next amino acid–tRNA complex
at the A site. paired to the codons. arrived amino acid. is approaching the ribosome.

71
 Elongation
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Met Met
asp
Met Met Ser Thr
Ser
peptide tRNA
Ser bond Ser Ala
Ala
Ala C U G Ala Trp Trp
peptide
Trp anticodon Trp Val bond Val
Val Val Asp
Asp Asp

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

3
5 3 5 5 3 5 3

1 A tRNA–amino acid 2 Two tRNAs can be at a 3 Peptide bond formation 4 The ribosome moves forward; the
approaches the ribosome at one time; attaches the peptide “empty” tRNA exits from the E site;
ribosome and binds the anticodons are chain to the newly the next amino acid–tRNA complex
at the A site. paired to the codons. arrived amino acid. is approaching the ribosome.

Elongation

72
 Second Step: Translation
• Termination:
 Previous tRNA moves to the P site
 Spent tRNA moves to the E site and exits
 Ribosome reads the STOP codon at the end of the
mRNA
• UAA, UAG, or UGA
• Does not code for an amino acid
 A protein called a release factor binds to the stop
codon and cleaves the polypeptide from the last tRNA
 The ribosome releases the mRNA and dissociates
into subunits
 The same mRNA may be read by another ribosome
73
 Termination
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Trp
release factor
Val

Trp

Val
U U
A A U G A

5′ stop codon 3′

The ribosome comes to a stop

codon on the mRNA. A release
factor binds to the site.

3′

5′

The release factor hydrolyzes the bond

between the last tRNA at the P site and
the polypeptide, releasing them. The
ribosomal subunits dissociate.

Termination 74
Summary of Protein Synthesis in Eukaryotes
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

TRANSCRIPTION 1. DNA in nucleus serves TRANSLATION

as a template for mRNA.

DNA 3. mRNA moves into

2. mRNA is processed large and small cytoplasm and
5 becomes associated
before leaving the nucleus.
mRNA ribosomal subunits
with ribosomes.
introns
pre-mRNA
3
mRNA amino 4. tRNAs with
acids
nuclear pore anticodons carry
amino acids
peptide to mRNA.

ribosome tRNA
U A C
A UG 3
5 anticodon

codon
5. During initiation, anticodon-codon
complementary base pairing begins
8. During termination, a
as the ribosomal subunits come
together at a start codon. ribosome reaches a stop
CCC UGG UU U
codon; mRNA and
5 GGG AC C AA A G ribosomal subunits
3 disband.

6. During elongation, polypeptide

synthesis takes place one
amino acid at a time.
7. Ribosome attaches to rough
ER. Polypeptide enters lumen,
where it folds and is
modified.

75
 12.6 Structure of the
Eukaryotic Chromosome
• Each chromosome contains a single linear DNA
molecule, but is composed of more than 50%
protein.
• Some of these proteins are concerned with DNA
and RNA synthesis
• Histones play primarily a structural role
 The most abundant proteins in chromosomes
 Five primary types of histone molecules
 Responsible for packaging the DNA
• The DNA double helix is wound at intervals around a core of
eight histone molecules (called a nucleosome)
• Nucleosomes are joined by “linker” DNA.
76
Structure of Eukaryotic Chromosomes Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

2nm

DNA
double helix
11 nm
1. Wrapping of DNA
a. Nucleosomes (“beads on a string”) around histone proteins.
histones
nucleosome

histone H1 2. Formation of a three-dimensional

zigzag structure via histone H1
and other DNA-binding proteins.
b. 30-nm fiber
30 nm

300 nm 3. Loose coiling into radial loops .

c. Radial loop domains

euchromatin

4. Tight compaction of radial

700 nm loops to form heterochromatin.

d. Heterochromatin

5. Metaphase chromosome forms

with the help of a protein
scaffold.
1,400 nm

e. Metaphase chromosome 77
a: © Ada L. Olins and Donald E. Olins/Biological Photo Service; b: Courtesy Dr. Jerome Rattner, Cell Biology and Anatomy, University of Calgary; c: Courtesy of Ulrich Laemmli and J.R. Paulson, Dept. of
Molecular Biology, University of Geneva, Switzerland; d: © Peter Engelhardt/Department of Pathology and Virology, Haartman Institute/Centre of Excellence in Computational Complex Systems Research,
Biomedical Engineering and Computational Science, Faculty of Information and Natural Sciences, Helsinki University of Technology, Helsinki, Finland