Chapter

10
DNA Structure
and Analysis
Lecture Presentation by
Dr. Cindy Malone,
California State University Northridge
2015 Pearson Education, Inc.

Section 10.1: Genetic Material

Genetic material
Information contained in genes that gets passed
onto new generation
Source of variability among organisms

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Section 10.1: Criteria for Genetic Material

To serve as genetic material, molecule must
be able to:
Replicate
Store information
Express information
Allow variation by mutation

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The Central Dogma

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Figure 10-1

Section 10.2: Protein as Genetic Material

In the 1940s, geneticists favored proteins as
genetic material
Proteins and nucleic acids were major candidates
for genetic material
Proteins were diverse and abundant in cells

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Section 10.2: Tetranucleotide Hypothesis

Tetranucleotide hypothesis
DNA contains equal amounts of four nucleotides
Postulated identical groups and repeats of four
components was basis for DNA structure
Lack of chemical diversity in DNA suggested it
could not store extensive genetic information
Proteins favored as genetic material

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10.3 Evidence Favoring DNA as the Genetic

Material Was First Obtained during the
Study of Bacteria and Bacteriophages

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Section 10.3: Avery, MacLeod, and McCarty

Avery, MacLeod, and McCarty
1944 publication on chemical nature of
transforming principle in bacteria
First direct experimental proof that DNA is
biomolecule responsible for heredity

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Section 10.3: Griffiths Transformation

Experiment
Griffith (1927)
Provided foundation for Avery, MacLeod, and
McCartys research
Showed avirulent strains of Diplococcus
pneumoniae could be transformed to virulence
(Figure 10-2)
Speculated transforming principle could be part
of polysaccharide capsule or compound required
for capsule synthesis

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 2015 Pearson Education, Inc.

Figure 10-2

Section 10.3: Transforming Principle is DNA

Avery, MacLeod, and McCarty
1944 published results of their experiment
DNase (deoxyribonuclease) utilized to destroy
transforming activity
Demonstrated transforming principle was DNA,
not protein (Figure 10-3)

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 2015 Pearson Education, Inc.

Figure 10-3

Section 10.3: Hershey and Chase

Hershey and Chase (1952)
Used Escherichia coli and bacteriophage T2
Demonstrated that DNA, not protein, is the
genetic material
Used radioisotopes 32P and 35S
Demonstrated DNA enters bacterial cell during
infection an directs viral reproduction
(Figures 10-4 and 10-5)

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 2015 Pearson Education, Inc.

Figure 10-4

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Figure 10-5

10.4 Indirect and Direct Evidence Supports

the Concept That DNA Is the Genetic
Material in Eukaryotes

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Section 10.4: Indirect Evidence of DNA as

Genetic Material
Indirect evidence: Distribution of DNA
Close correlation between gametes and diploids
in amount of DNA and number of chromosome
sets
No such correlation between gametes and
diploids for proteins

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Section 10.4: Indirect Evidence: Mutagenesis

Indirect evidence: Mutagenesis
UV light: Most mutagenic at wavelength
260 nm (action spectrum)
DNA absorbs UV at 260 nm
Protein absorbs UV at 280 nm
A wavelength at which no significant mutagenic effects
are observed

Molecule serving as genetic material expected to

absorb at mutagenic wavelength (Figure 10-6)

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 2015 Pearson Education, Inc.

Figure 10-6

Section 10.4: Direct Evidence: Recombinant

DNA Studies
Recombinant DNA technology
Segments of eukaryotic DNA corresponding to
specific genes isolated and spliced into the
bacterial DNA
Complex inserted into bacterial cell and monitored

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 2015 Pearson Education, Inc.

Section 10.4: Direct Evidence

Eukaryotic DNA now functional in bacterial
cell
Eukaryotic gene product in bacteria containing
eukaryotic gene provides direct evidence: DNA is
present and functional in bacterial cell
Example: Insulin and interferon production by
bacteria

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 2015 Pearson Education, Inc.

Section 10.4: Direct Evidence: Transgenic

Animals
Transgenic animal example
Human DNA microinjected into fertilized mouse
egg
DNA encoded human -globin gene
Now present and expressed in mouse and
transmitted to progeny

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10.5 RNA Serves as the Genetic

Material in Some Viruses

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Section 10.5: RNA as Genetic Material in

Some Viruses
Some viruses have RNA core, not DNA
TMV: Tobacco mosaic virus (1956)
Demonstrated RNA serves as genetic material for
these viruses

RNA replicase
Enzyme isolated from E. coli (1965, Pace and
Spiegelman)
Replication of the viral RNA is dependent on RNA
replicase
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Section 10.5: Retroviruses and Reverse

Transcriptase
Retroviruses
Replicate unusually
RNA serves as template for DNA synthesis
Complementary synthesis of DNA by RNAdependent DNA polymerase reverse
transcriptase

Reverse transcriptase
RNA-dependent DNA polymerase enzyme

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 2015 Pearson Education, Inc.

10.6 Knowledge of Nucleic Acid Chemistry

Is Essential to the Understanding of DNA
Structure

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Section 10.6: Nucleotides

Nucleotides
DNA is nucleic acid
Nucleotides: Building blocks of nucleic acid
Nucleotides are building blocks of DNA

Nucleotides consist of:

Nitrogenous base (two kinds)
Pentose sugar
Phosphate group
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Section 10.6: Nitrogenous Bases

Two kinds of nitrogenous bases
Purines (nine-member ring)
Adenine (A)
Guanine (G)

Pyrimidines (six-member ring)

Cytosine (C)
Thymine (T)
Uracil (U)

Figure 10-7

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 2015 Pearson Education, Inc.

Figure 10-7

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Figure 10-7a

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Figure 10-7b

Section 10.6: The Bases of DNA and RNA

DNA
Bases are A, C, T, G

RNA
Bases are A, C, U, G

Only DNA contains T

Only RNA contains U

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Section 10.6: Ribose and Deoxyribose

RNA contains ribose sugar
DNA contains deoxyribose
Deoxy (without an oxygen) (Figure 10-7b)

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 2015 Pearson Education, Inc.

Figure 10-7

Section 10.6: Nucleosides and Nucleotides

Nucleoside
Contains nitrogenous base and pentose sugar
Molecule is composed of purine or pyrimidine
base and ribose or deoxyribose sugar

Nucleotide
Nucleoside with phosphate group added
(Figure 10-8)

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 2015 Pearson Education, Inc.

Figure 10-8

Section 10.6: Mono-, Di-, and Triphosphates

Nucleoside monophosphates (NMP)
A nucleotide

Nucleoside diphosphates (NDP)

Nucleotide with addition of two phosphate groups

Nucleoside triphosphates (NTP)

Nucleotide with addition of three phosphate
groups

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Section 10.6: Triphosphates

Triphosphates
Serve as precursor molecule during nucleic acid
synthesis

ATP and GTP: Adenosine triphosphate and

guanine triphosphate
Large amount of energy involved in
adding/removing terminal phosphate group

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 2015 Pearson Education, Inc.

Figure 10-9

Section 10.6: Phosphodiester Bonds

Phosphodiester bonds
Nucleotides are linked by phosphodiester
bonds between phosphate group at C-5 position
and OH group on C-3 position (Figure 10-10)

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 2015 Pearson Education, Inc.

Figure 10-10

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Figure 10-10a

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Figure 10-10b

10.7 The Structure of DNA Holds the Key to

Understanding Its Function

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Section 10.7: Structure of DNA

Watson and Crick, 1953
Proposed structure of DNA as a double helix

Chargaff (19491953)
Proposed base composition
Amount of A is proportional to T
Amount of C is proportional to G
Percentage of C G does not equal percentage
of A T (Table 10.3)

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Section 10.7: X-Ray Diffraction

Base composition analysis (Chargaff) and Xray diffraction provided crucial data to Watson
and Crick
X-ray diffraction
Studies by Rosalind Franklin (19501953)
showed DNA had a 3.4-angstrom periodicity,
characteristic of helical structure (Figure 10-11)

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 2015 Pearson Education, Inc.

Figure 10-11

Section 10.7: Structure of DNA

Watson and Crick model (1953)
Proposed DNA as:
Double helix
Two anti-parallel strands connected by base
pairing
Stacked nitrogenous bases (Figure 10-12)

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 2015 Pearson Education, Inc.

Figure 10-12

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Figure 10-12a

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Figure 10-12b

Section 10.7: Base Pairing Hydrogen Bonds

Chemical affinity produces hydrogen bonds in
pair of bases
A-T and G-C base pairing provides
complementarity of two strands and chemical
stability to the helix
A-T: Double bond
G-C: Triple bond (Figure 10-14)

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 2015 Pearson Education, Inc.

Figure 10-14

Section 10.7: Semiconservative Model of

Replication
Watson and Crick: Semiconservative model
Storage of genetic information in sequence of
bases
Mutations or genetic changes that could result in
alteration of bases

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10.8 Alternative Forms of DNA Exist

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Section 10.8: Alternative Forms of DNA

Watson-Crick DNA model of B-DNA
Seen under aqueous, low-salt conditions

Alternative forms of DNA

Different conformations of DNA observed under
different conditions of isolation

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10.9 The Structure of RNA Is Chemically

Similar to DNA, but Single Stranded

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Section 10.9: Structure of RNA

RNA
Sugar ribose replaces deoxyribose of DNA
Nitrogenous base uracil replaces thymine of DNA
Most are single-stranded (ss)
Exception: Animal viruses have double-stranded
helices

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Section 10.9: Three Classes of RNAs

Three classes of cellular RNAs (function during
gene expression)
Messenger RNA (mRNA)
Ribosomal RNA (rRNA)
Transfer RNA (tRNA)
(Table 10.4)

Originate as complementary copies of one of two

DNA strands during transcription

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 2015 Pearson Education, Inc.

Table 10.4

Section 10.9: Three Major Classes of RNAs

rRNAs: Ribosomal RNAs
Structural components of ribosomes for protein
synthesis

mRNAs: Messenger RNAs

Template for protein synthesis
Carry genetic information from gene to ribosome

tRNAs: Transfer RNAs

Carry amino acids for protein synthesis

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Section 10.9: Unique RNAs in Eukaryotes

Unique RNAs
Telomerase RNA and RNA primers
Involved in DNA replication at chromosome ends

snRNA: Small nuclear RNA

Process mRNAs

Antisense RNA, microRNA, siRNA

Involved in gene regulation

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10.10 Many Analytical Techniques Have

Been Useful during the Investigation
of DNA and RNA

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Section 10.1: Analytical Techniques

Analytical techniques useful during DNA and
RNA investigation
Absorption of UV light
Electrophoresis of nucleic acids

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Section 10.10: Absorption of UV Light

Nucleic acids
Absorb UV light strongly at 254260 nm due to
interaction between UV light and ring systems of
the bases (Figure 10-16)
UV light used in localization, isolation, and
characterization
Use of UV critical to isolation of nucleic acids
following separation

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Section 10.10: Electrophoresis

Nucleic acid electrophoresis
Separates DNA and RNA fragments by size
Smaller fragments migrate through gel at faster
rate than large fragments (Figure 10-19)
Agarose gel
Porous matrix restricts migration of larger molecules
more than it restricts smaller ones

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 2015 Pearson Education, Inc.

Figure 10-19