Chapter 16: The Molecular Basis of

Inheritance

Campbell Biology, 12th Edition

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
• This DNA program directs the development of
biochemical, anatomical, physiological, and (to
some extent) behavioral traits
Concept 16.1: DNA is the genetic material

• Early in the 20th century, the identification of the

molecules of inheritance loomed as a major challenge to
biologists
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 and one harmless.
• When he mixed heat-killed remains of the
pathogenic strain with living cells of the harmless
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.
 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
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.
 Phage
head

Tail
sheath

Tail fiber

DNA

100 nm
Bacterial
cell
 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.
 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

• 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.
(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.
 5 end
C G
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).
•At first, Watson and Crick thought the bases
paired like with like (A with A, and so on), but
such pairings did not result in a uniform width.

•Instead, pairing a purine with a pyrimidine

resulted in a uniform width consistent with the
X-ray data.
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.
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.
 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
• Watson and Crick’s semiconservative model of
replication predicts that when a double helix
replicates, each daughter molecule will have one old
strand (derived or “conserved” from the parent
molecule) and one newly made strand.

• Competing models were the conservative model (the

two parent strands rejoin) and the dispersive model
(each strand is a mix of old and new).
 Parent First Second
cell replication replication

(a) Conservative
model

(b) Semiconservative
model

(c) Dispersive model

• Experiments by Matthew Meselson and Franklin
Stahl supported the semiconservative model.

• They labeled the nucleotides of the old strands with

a heavy isotope of nitrogen, while any new
nucleotides were labeled with a lighter isotope.
EXPERIMENT
1 Bacteria 2 Bacteria
cultured in transferred to
medium with medium with
15N (heavy 14N (lighter

isotope) isotope)

RESULTS
3 DNA sample 4 DNA sample Less
centrifuged centrifuged dense
after first after second
replication replication More
dense
CONCLUSION
Predictions: First replication Second replication

Conservative
model

Semiconservative
model

Dispersive
model
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.
(a) Origin of replication in an E. coli cell
Origin of
replication Parental (template) strand

Daughter (new) strand

Double-
stranded Replication fork
DNA molecule Replication
bubble

Two
daughter
DNA molecules

0.5 m
(b) Origins of replication in a eukaryotic cell
Double-stranded
Origin of replication DNA molecule

Parental (template) Daughter (new)

strand strand

Bubble Replication fork

Two daughter DNA molecules

0.25 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.
• 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.
 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 PiOH
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.
• 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.
 3
5 3
Template
strand 5
3 RNA primer
for fragment 1
5
1 3
5

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

2
1 3
5 5
3
2
1 3
5
Overall direction of replication
 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
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.
 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.
5 3

3 5
Nuclease

5 3

3 5

DNA
polymerase

5 3

3 5
DNA
ligase

5 3

3 5