DNA
The Genetic Material
Functions of the Genetic Material
The genetic material must replicate, control the
growth and development of the organism, and
allow the organism to adapt to changes in the
environment.
Genotypic Function: Replication
Phenotypic Function: Gene Expression
Evolutionary Function: Mutation (Gene modifications)
1928 - Frederick Griffith’s Transformation Experiments
• Two strains of Streptococcus pneumoniae
– S Smooth
• Secrete a polysaccharide capsule
• Produce smooth colonies on solid media
• virulent
– R Rough
• Unable to secrete a capsule
• Produce colonies with a rough appearance
• avirulent
• Two coat types
– II
– III
• Four possible phenotypes
– IIS or IIIS or IIR or IIIR
Figure 9.2
9-8
SUMMARY 5
Griffith’s Conclusions
Something from the dead type IIIS transformed
type IIR into type IIIS
Called this process transformation
The unknown substance was termed the
transforming principle
9-9
Sia and Dawson’s experiment (in vitro) in 1931
1944 - Avery, MacLeod & McCarty
Identify the Genetic Material
• Griffith’s transforming principle was the
genetic material
• Transformation assay to identify actual
biomolecule
• Major constituents - DNA, RNA, proteins,
carbohydrates, & lipids
• Made cell extracts from type IIIS cells
containing each of these macromolecules
9-10
Avery’s Experiments
Mixed each extract with type IIR cells to test for transformation
Only extract containing purified DNA transformed type IIR to
type IIIS
Verify that DNA, not RNA or protein, is the genetic material
Figure 9.3 9-11
The Hershey and Chase experiment:
• 1952- Hershey & Chase Confirm DNA is
Genetic Material Using Phage T2
• Used radioisotopes to distinguish DNA from
proteins
– 32P labels DNA specifically
– 35S labels protein specifically
• Infect non-radioactive E. coli with radioactively-
labeled phages
• Remove phage coats from cells
• Is 32P or 35S inside bacteria?
Life cycle of
phage T2
Figure 9.5
Hershey & Chase Experiment
S
Hershey & Chase Experiment
Summary of Hershey & Chase Experiment
The Discovery of RNA as Viral Genetic Material
All known cellular organisms have DNA
as their genetic material.
Some viruses, however, use RNA
instead.
Gierer and Schramm
Experiment (1956)
Tobacco mosaic virus (TMV) is composed of RNA and protein; it contains
no DNA.
In 1956 Gierer and Schramm showed that when purified RNA from TMV
is applied directly to tobacco leaves, they develop mosaic disease.
Pretreating the purified RNA with RNase destroys its ability to cause TMV
lesions .
Typical tobacco mosaic
virus (TMV) particle
Fraenkel-Conrat & co-workers Experiment
(1957)
• In 1957 Fraenkel-Conrat & co-workers showed that in
TMV infections with viruses containing RNA from one
strain and protein from another, the progeny viruses
were always of the type specified by the RNA, not by the
protein.
Demonstration that RNA is the genetic material in
tobacco mosaic virus (TMV)
Final conclusion
• The genetic information of most living
organisms is stored in deoxyribonucleic acid
(DNA).
• In some viruses, the genetic information is
present in ribonucleic acid (RNA).
The Composition of DNA and RNA
DNA and RNA are polymers composed of
monomers called nucleotides.
Each nucleotide has three parts:
a. A pentose (5-carbon) sugar.
b. A nitrogenous base.
c. A phosphate group.
The pentose sugar in RNA is ribose, and in DNA it’s
deoxyribose. The only difference is at the 2’ position, where
RNA has a hydroxyl (OH) group, while DNA has only a
hydrogen.
Structures of deoxyribose and ribose in DNA and RNA
There are two classes of nitrogenous bases
a. Purines (double-ring, nine-membered structures)
include adenine (A) and guanine (G).
b. Pyrimidines (one-ring, six-membered structures)
include cytosine (C), thymine (T) in DNA and uracil
(U) in RNA.
Structures of the nitrogenous bases in DNA and RNA
Why Thymine not Uracil in DNA?
First Reason CH3
• Despite uracil’s tendency to pair with adenine, it can
also pair with any other base, including itself.
• By adding a methyl group (which is hydrophobic) and
turning it into thymine, its position is reorganized in
the double-helix, not allowing those wrong pairings to
happen.
Second Reason
• Cytosine can deaminate to produce uracil. The only
difference between them is the change from an O in uracil
to an NH2 in cytosine.
• The problem is that, if uracil were a component of DNA,
the repair systems would not be able to distinguish
original uracil from uracil originated by deamination of
cytosine.
• So using thymine instead makes it way easier and more
stable, as any uracil inside DNA must come from a cytosine
and so it can be replaced by a new cytosine.
Watson & Crick’s DNA Model
Two additional sources of data
assisted Watson and Crick with
their model:
a. Erwin Chargaff’s ratios obtained for
DNA derived from a variety of
sources showed that the amount of
purine always equals the amount of
pyrimidine, and further, that the
amount of G equals C, and the
amount of A equals T.
b. Rosalind Franklin’s X ray diffraction
images of DNA showed a helical
structure with regularities at 0.34 nm
and 3.4 nm along the axis of the
molecule (Figure 8.9).
Wilkins & Franklin’s Photographs
• X-ray diffraction to study
the structures of
molecules
• 1952 Wilkins & Franklin
developed high-quality X-
ray diffraction
photographs of strands
of DNA which suggested
that the DNA resembled
a tightly coiled helix and
was composed of two or
three chains of
nucleotides
The Double Helix
Phosphodiester
[C-O-P-O-C]
Polynucleotide chain
Complementary and Antiparallel
DNA Structure
• Complementary Base Pairs (A with T, G with C)
• Anti-parallel Strands
• Right-handed double helix (B-DNA)
Different DNA Structures
X ray diffraction studies show that DNA can exist in different
forms
a. A-DNA is the dehydrated form, and so it is not usually found in
cells. It is a right-handed helix with 10.9 bp/turn. A-DNA has a
deep and narrow major groove, and a wide and shallow minor
groove.
b. B-DNA is the hydrated form of DNA, the kind normally found in
cells. It is also a right-handed helix, with only 10.0 bp/turn. B-
DNA has a wide major groove and a narrow minor groove, and
its major and minor grooves are of about the same depth.
c. Z-DNA is a left-handed helix with a zigzag sugar-phosphate
backbone that gives it its name. It has 12.0 bp/turn in helix. Z-
DNA has a deep minor groove, and a very shallow major groove.
Its existence in living cells has not been proven.
DNA replication DNA-RNA duplexes
Elevated content of C:G
THE GENETIC CODE
Most genetic code tables designate the codons for
amino acids as mRNA sequences. Important features of
the genetic code include:
• Each codon consists of three bases (triplet). There are
64 codons. They are all written in the 5' to 3' direction.
• 61 codons code for amino acids. The other three
(UAA, UGA, UAG) are stop codons (or nonsense
codons) that terminate translation.
• There is one start codon (initiation codon), AUG,
coding for methionine. Protein synthesis begins with
methionine (Met) in eukaryotes, and formyl
methionine (fmet) in prokaryotes.
• The code is unambiguous. Each codon specifies no
more than one amino acid.
• The code is degenerate. More than one codon can
specify a single amino acid.
• All amino acids, except Met and tryptophan (Trp), have
more than one codon.
• For those amino acids having more than one codon, the
first two bases in the codon are usually the same. The
base in the third position often varies.
• The code is almost universal (the same in all
organisms). Some minor exceptions to this occur in
mitochondria and some organisms.
• The code is commaless (contiguous). There are no
spacers or "commas" between codons on an mRNA.
• Neighboring codons on a message are non-overlapping.
