LECTURE PRESENTATIONS

For CAMPBELL BIOLOGY, NINTH EDITION

Jane B. Reece, Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, Robert B. Jackson

Chapter 16

The Molecular Basis of

Inheritance

Lectures by
Erin Barley
Kathleen Fitzpatrick

© 2011 Pearson Education, Inc.

Overview: Life’s Operating Instructions
• In 1953, James Watson and Francis Crick
introduced an elegant double-helical model for
the structure of deoxyribonucleic acid, or DNA
• DNA, the substance of inheritance, is the most
celebrated molecule of our time
• Hereditary information is encoded in DNA and
reproduced in all cells of the body
Figure 16.1
The Search for the Genetic Material:
Scientific Inquiry
• When T. H. Morgan’s group showed that genes
are located on chromosomes, the two components
of chromosomes—DNA and protein—became
candidates for the genetic material
• The role of DNA in heredity was first discovered by
studying bacteria and the viruses that infect them
Evidence That DNA Can Transform Bacteria
• The discovery of the genetic role of DNA began
with research by Frederick Griffith in 1928
• Griffith worked with two strains of a bacterium, one
pathogenic (S) and one harmless (R)
• When he mixed heat-killed remains of the
pathogenic (S) strain with living cells of the
harmless (R) strain, some living cells became
pathogenic
• He called this phenomenon transformation, now
defined as a change in genotype and phenotype
due to assimilation of foreign DNA
Figure 16.2
EXPERIMENT Mixture of
Heat-killed heat-killed
Living S cells Living R cells S cells S cells and
(control) (control) (control) living R cells

RESULTS

Mouse dies Mouse healthy Mouse healthy Mouse dies

Living S cells
• In 1944, Oswald Avery, Maclyn McCarty, and Colin
MacLeod announced that the transforming
substance was DNA
• Their conclusion was based on experimental
evidence that only DNA worked in transforming
harmless bacteria into pathogenic bacteria
Evidence That Viral DNA Can Program Cells
• More evidence for DNA as the genetic material
came from studies of viruses that infect bacteria
• Such viruses, called bacteriophages (or phages),
are widely used in molecular genetics research
Figure 16.3

Phage
head

Tail
sheath

Tail fiber

DNA

100 nm
Bacterial
cell
• In 1952, Alfred Hershey and Martha Chase
performed experiments showing that DNA is the
genetic material of a phage known as T2
Figure 16.4-3

EXPERIMENT
Radioactive Empty
protein protein
shell Radioactivity
Phage (phage protein)
in liquid
Bacterial cell

Batch 1:
Radioactive DNA
sulfur Phage
(35S) DNA

Centrifuge

Radioactive Pellet (bacterial

DNA cells and contents)

Batch 2:
Radioactive
phosphorus
(32P)

Centrifuge
Radioactivity
Pellet (phage DNA)
in pellet
Additional Evidence That DNA Is the
Genetic Material
• It was known that DNA is a polymer of nucleotides,
each consisting of a nitrogenous base, a sugar,
and a phosphate group
• In 1950, Erwin Chargaff reported that DNA
composition varies from one species to the next
• This evidence of diversity made DNA a more
credible candidate for the genetic material
• Two findings became known as Chargaff’s rules
– The base composition of DNA varies between species
– In any species the number of A and T bases are equal
and the number of G and C bases are equal
• The basis for these rules was not understood until
the discovery of the double helix
Figure 16.5
Sugar–phosphate Nitrogenous bases
backbone
5 end

Thymine (T)

Adenine (A)

Cytosine (C)

Phosphate
Guanine (G)

Sugar
(deoxyribose)
DNA Nitrogenous base
nucleotide 3 end
Building a Structural Model of DNA:
Scientific Inquiry
• After DNA was accepted as the genetic material,
the challenge was to determine how its structure
accounts for its role in heredity
• Maurice Wilkins and Rosalind Franklin were using
a technique called X-ray crystallography to study
molecular structure
• Franklin produced a picture of the DNA molecule
using this technique
Figure 16.6

(a) Rosalind Franklin (b) Franklin’s X-ray diffraction

photograph of DNA
• Franklin’s X-ray crystallographic images of DNA
enabled Watson to deduce that DNA was helical
• The X-ray images also enabled Watson to deduce
the width of the helix and the spacing of the
nitrogenous bases
• The pattern in the photo suggested that the DNA
molecule was made up of two strands, forming a
double helix
Figure 16.7

G
5 end
C
C G Hydrogen bond
3 end
G C
G C T A

3.4 nm
T A

G C G C
C G

A T

1 nm C G
T A
C G
G C
C G A T

A T 3 end
A T
0.34 nm
T A 5 end

(a) Key features of (b) Partial chemical structure (c) Space-filling

DNA structure model
• Watson and Crick built models of a double helix to
conform to the X-rays and chemistry of DNA
• Franklin had concluded that there were two outer
sugar-phosphate backbones, with the nitrogenous
bases paired in the molecule’s interior
• Watson built a model in which the backbones were
antiparallel (their subunits run in opposite
directions)
Figure 16.UN01

Purine + purine: too wide

Pyrimidine + pyrimidine: too narrow

Purine + pyrimidine: width

consistent with X-ray data
• Watson and Crick reasoned that the pairing was
more specific, dictated by the base structures
• They determined that adenine (A) paired only with
thymine (T), and guanine (G) paired only with
cytosine (C)
• The Watson-Crick model explains Chargaff’s rules:
in any organism the amount of A = T, and the
amount of G = C
Figure 16.8

Sugar
Sugar
Adenine (A) Thymine (T)

Sugar
Sugar

Guanine (G) Cytosine (C)

Concept 16.2: Many proteins work
together in DNA replication and repair
• The relationship between structure and function is
manifest in the double helix
• Watson and Crick noted that the specific base
pairing suggested a possible copying mechanism
for genetic material
The Basic Principle: Base Pairing to a
Template Strand
• Since the two strands of DNA are complementary,
each strand acts as a template for building a new
strand in replication
• In DNA replication, the parent molecule unwinds,
and two new daughter strands are built based on
base-pairing rules
Figure 16.9-3

A T A T A T A T
C G C G C G C G
T A T A T A T A
A T A T A T A T
G C G C G C G C

(a) Parent molecule (b) Separation of (c) “Daughter” DNA molecules,

strands each consisting of one
parental strand and one
new strand
DNA Replication: A Closer Look
• The copying of DNA is remarkable in its speed and
accuracy
• More than a dozen enzymes and other proteins
participate in DNA replication
Getting Started
• Replication begins at particular sites called
origins of replication, where the two DNA
strands are separated, opening up a replication
“bubble”
• A eukaryotic chromosome may have hundreds or
even thousands of origins of replication
• Replication proceeds in both directions from each
origin, until the entire molecule is copied
Figure 16.12
(a) Origin of replication in an E. coli cell (b) Origins of replication in a eukaryotic cell
Origin of Double-stranded
Parental (template) strand Origin of replication DNA molecule
replication
Daughter (new)
strand Parental (template) Daughter (new)
strand strand
Replication
Double- fork
stranded
DNA molecule Replication
bubble
Bubble Replication fork

Two daughter
DNA molecules

Two daughter DNA molecules

0.25 m
0.5 m
• At the end of each replication bubble is a
replication fork, a Y-shaped region where new
DNA strands are elongating
• Helicases are enzymes that untwist the double
helix at the replication forks
• Single-strand binding proteins bind to and
stabilize single-stranded DNA
• Topoisomerase corrects “overwinding” ahead of
replication forks by breaking, swiveling, and
rejoining DNA strands
Figure 16.13

Primase

3
Topoisomerase
5 RNA
3 primer
5
3

Helicase

5
Single-strand binding
proteins
• DNA polymerases cannot initiate synthesis of a
polynucleotide; they can only add nucleotides to
the 3 end
• The initial nucleotide strand is a short RNA
primer
• An enzyme called primase can start an RNA
chain from scratch and adds RNA nucleotides one
at a time using the parental DNA as a template
• The primer is short (5–10 nucleotides long), and
the 3 end serves as the starting point for the new
DNA strand
Synthesizing a New DNA Strand
• Enzymes called DNA polymerases catalyze the
elongation of new DNA at a replication fork
• Most DNA polymerases require a primer and a
DNA template strand
• The rate of elongation is about 500 nucleotides
per second in bacteria and 50 per second in
human cells
• Each nucleotide that is added to a growing DNA
strand is a nucleoside triphosphate
• dATP supplies adenine to DNA and is similar to
the ATP of energy metabolism
• The difference is in their sugars: dATP has
deoxyribose while ATP has ribose
• As each monomer of dATP joins the DNA strand, it
loses two phosphate groups as a molecule of
pyrophosphate
Figure 16.14

New strand Template strand

5 3 5 3

Sugar A T A T
Phosphate Base

C G C G

G C G C
DNA
OH
polymerase
3 A T A
P Pi OH
C Pyrophosphate 3 C

Nucleoside 2Pi
triphosphate 5 5
Antiparallel Elongation
• The antiparallel structure of the double helix
affects replication
• DNA polymerases add nucleotides only to the free
3 end of a growing strand; therefore, a new DNA
strand can elongate only in the 5 to 3 direction

• Along one template strand of DNA, the DNA

polymerase synthesizes a leading strand
continuously, moving toward the replication fork
Figure 16.15
Overview
Leading
strand Origin of replication Lagging
strand

Primer

Lagging Leading
strand strand Origin of
Overall directions replication
of replication

3
5

5 RNA primer
3
3 Sliding clamp

DNA pol III

Parental DNA 5
3
5

5
3
3

5
• To elongate the other new strand, called the
lagging strand, DNA polymerase must work in the
direction away from the replication fork
• The lagging strand is synthesized as a series of
segments called Okazaki fragments, which are
joined together by DNA ligase
Figure 16.16
3
Overview
5 3 Leading Origin of replication Lagging
Template
strand strand strand
RNA primer 5
for fragment 1 Lagging strand
3
2
1
5 Leading
1 3 strand
Overall directions
5 of replication

3
Okazaki
fragment 1
5
1
RNA primer 3
for fragment 2 5
5 Okazaki
3 fragment 2
2

1 3
5 5
3

2
1 3
5
5
3

2
1
3
5
Overall direction of replication
Figure 16.17

Overview
Leading Origin of
replication Lagging
strand strand

Leading
Lagging strand
strand Overall directions
Leading strand of replication

5 DNA pol III

3 Primer
Primase
3 5
3
Parental DNA pol III Lagging strand
DNA 5
4 DNA pol I DNA ligase
35
3 2 1 3

5
Figure 16.17a
Overview
Leading Origin of
replication Lagging
strand strand

Leading
Lagging strand
strand Overall directions
of replication

Leading strand

5 DNA pol III

3 Primer
Primase
3 5
3
Parental
DNA
Figure 16.17b
Overview
Leading Origin of
replication Lagging
strand strand

Leading
Lagging strand
strand Overall directions
Leading strand of replication

Primer

DNA pol III Lagging strand

5
4 DNA pol I DNA ligase
35
3 3 2 1 3

5
The DNA Replication Complex
• The proteins that participate in DNA replication
form a large complex, a “DNA replication machine”
• The DNA replication machine may be stationary
during the replication process
• Recent studies support a model in which DNA
polymerase molecules “reel in” parental DNA and
“extrude” newly made daughter DNA molecules
Figure 16.18

DNA pol III

Parental DNA Leading strand
5
5 3 3

3
3 5 5

Connecting Helicase
protein

3 5 Lagging
DNA strand
Lagging strand template
pol III 5 3
Proofreading and Repairing DNA
• DNA polymerases proofread newly made DNA,
replacing any incorrect nucleotides
• In mismatch repair of DNA, repair enzymes
correct errors in base pairing
• DNA can be damaged by exposure to harmful
chemical or physical agents such as cigarette
smoke and X-rays; it can also undergo
spontaneous changes
• In nucleotide excision repair, a nuclease cuts
out and replaces damaged stretches of DNA
Figure 16.19
5 3

3 5
Nuclease

5 3

3 5

DNA
polymerase

5 3

3 5
DNA
ligase

5 3

3 5
Evolutionary Significance of Altered DNA
Nucleotides
• Error rate after proofreading repair is low but not
zero
• Sequence changes may become permanent and
can be passed on to the next generation
• These changes (mutations) are the source of the
genetic variation upon which natural selection
operates
Replicating the Ends of DNA Molecules
• Limitations of DNA polymerase create problems
for the linear DNA of eukaryotic chromosomes
• The usual replication machinery provides no way
to complete the 5 ends, so repeated rounds of
replication produce shorter DNA molecules with
uneven ends
• This is not a problem for prokaryotes, most of
which have circular chromosomes
Figure 16.20
5
Ends of parental Leading strand
DNA strands Lagging strand
3

Last fragment Next-to-last fragment

Lagging strand RNA primer

5
3
Parental strand
Removal of primers and
replacement with DNA
where a 3 end is available
5
3
Second round
of replication

5
New leading strand 3
New lagging strand 5
3
Further rounds
of replication

Shorter and shorter daughter molecules

• Eukaryotic chromosomal DNA molecules have
special nucleotide sequences at their ends called
telomeres
• Telomeres do not prevent the shortening of DNA
molecules, but they do postpone the erosion of
genes near the ends of DNA molecules
• It has been proposed that the shortening of
telomeres is connected to aging
Figure 16.21

1 m
• If chromosomes of germ cells became shorter in
every cell cycle, essential genes would eventually
be missing from the gametes they produce
• An enzyme called telomerase catalyzes the
lengthening of telomeres in germ cells
• The shortening of telomeres might protect cells
from cancerous growth by limiting the number of
cell divisions
• There is evidence of telomerase activity in cancer
cells, which may allow cancer cells to persist
Concept 16.3 A chromosome consists of a
DNA molecule packed together with proteins
• The bacterial chromosome is a double-stranded,
circular DNA molecule associated with a small
amount of protein
• Eukaryotic chromosomes have linear DNA
molecules associated with a large amount of
protein
• In a bacterium, the DNA is “supercoiled” and found
in a region of the cell called the nucleoid
• Chromatin, a complex of DNA and protein,
is found in the nucleus of eukaryotic cells
• Chromosomes fit into the nucleus through
an elaborate, multilevel system of packing
Figure 16.22a

Nucleosome
(10 nm in diameter)
DNA double helix
(2 nm in diameter)

H1
Histone
Histones tail
Nucleosomes, or “beads on
DNA, the double helix Histones a string” (10-nm fiber)
Figure 16.22b

Chromatid
(700 nm)

30-nm fiber

Loops Scaffold

300-nm fiber

30-nm fiber

Replicated
chromosome
(1,400 nm)
Looped domains
(300-nm fiber) Metaphase
chromosome
Figure 16.UN02

G C
A T
T A
Nitrogenous bases
G C
Sugar-phosphate backbone C G
A T
C G

Hydrogen bond
T A
Figure 16.UN03

DNA pol III synthesizes

leading strand continuously 3
5
Parental
DNA DNA pol III starts DNA
synthesis at 3 end of primer, Origin of
5 continues in 5 → 3 direction replication
3
5 Lagging strand synthesized
in short Okazaki fragments,
Helicase later joined by DNA ligase

Primase synthesizes 3
a short RNA primer 5
DNA pol I replaces the RNA
primer with DNA nucleotides