Download The Molecular Basis of Heredity

The Search for the Genetic
Material of Life
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What is a gene?
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Stable source of information
Ability to replicate accurately
Capable of change
The Search for the Molecular Basis of Heredity
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Search for genetic material---nucleic acid or protein/DNA
or RNA?
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Griffith’s Transformation Experiment
Avery’s Transformation Experiment
Hershey-Chase Bacteriophage Experiment
Tobacco Mosaic Virus (TMV) Experiment
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Nucleotides - composition and structure
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Double-helix model of DNA - Watson & Crick
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Original Source for portions of slide content:
http://mercury.bio.uaf.edu/~kevin_mccracken/genetics/lectures/chapter_02.ppt by
Kevin McCracken University of Alaska Fairbanks.
Timeline of events
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1890
Weismann - substance in the cell nuclei controls
development.
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1900
Chromosomes shown to contain hereditary
information, later shown to be composed of protein &
nucleic acids.
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1928
Griffith’s Transformation Experiment
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1944
Avery’s Transformation Experiment
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1953
Hershey-Chase Bacteriophage Experiment
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1953
Watson & Crick propose double-helix model of DNA
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1956
Gierer & Schramm/Fraenkel-Conrat & Singer
Demonstrate RNA is viral genetic material.
Frederick Griffith’s Transformation Experiment - 1928
“transforming principle” demonstrated with Streptococcus pneumoniae
Griffith hypothesized that the transforming agent was a “IIIS” protein.
Oswald T. Avery’s Transformation Experiment 1944
Determined that “IIIS” DNA was the genetic material
responsible for Griffith’s results (not RNA).
Hershey-Chase Bacteriophage Experiment - 1953
Bacteriophage =
Virus that
attacks bacteria
and replicates
by invading a
living cell and
using the cell’s
molecular
machinery.
Structure of T2
phage
DNA & protein
Life cycle of virulent T2 phage:
Hershey-Chase Bacteriophage Experiment - 1953
1.
2.
T2 bacteriophage is
composed of DNA and
proteins:
Set-up two replicates:
•
•
Label DNA with 32P
Label Protein with 35S
3.
Infected E. coli bacteria with
two types of labeled T2
4.
32P
is discovered within the
bacteria and progeny
phages, whereas 35S is not
found within the bacteria but
released with phage ghosts.
1969: Alfred Hershey
Gierer & Schramm Tobacco Mosaic Virus (TMV) Experiment –
1956 & Fraenkel-Conrat & Singer - 1957
• Used 2 viral strains to demonstrate RNA is the genetic material of TMV
Conclusions about these early
experiments:
•
Griffith 1928 & Avery 1944:
•
•
Hershey-Chase 1953:
•
•
DNA (not RNA) is transforming agent.
DNA (not protein) is the genetic material.
Gierer & Schramm 1956/Fraenkel-Conrat &
Singer 1957:
•
RNA (not protein) is genetic material of
some viruses.
Nucleotide = monomers that make up DNA and RNA (Figs. 2.9-10)
Three components
1. Pentose (5-carbon) sugar
DNA = deoxyribose
RNA = ribose
(compare 2’ carbons)
2. Nitrogenous base
Purines
Adenine
Guanine
Pyrimidines
Cytosine
Thymine (DNA)
Uracil (RNA)
3. Phosphate group attached to 5’ carbon
Nucleotides are linked by phosphodiester bonds
to form polynucleotides.
Phosphodiester bond
Covalent bond between the phosphate group (attached to
5’ carbon) of one nucleotide and the 3’ carbon of the
sugar of another nucleotide.
This bond is very strong, and for this reason DNA is
remarkably stable. DNA can be boiled and even
autoclaved without degrading!
5’ and 3’
The ends of the DNA or RNA chain are not the same. One
end of the chain has a 5’ carbon and the other end has a
3’ carbon.
5’ end
3’ end
James D. Watson & Francis H. Crick - 1953
Double Helix Model of DNA
Two sources of information:
1.
Base composition studies of Erwin Chargaff
•
indicated double-stranded DNA consists of ~50% purines
(A,G) and ~50% pyrimidines (T, C)
•
amount of A = amount of T and amount of G = amount of C
(Chargraff’s rules)
•
%GC content varies from organism to organism
Examples:
%A
%T
%G
%C
%GC
Homo sapiens
Zea mays
Drosophila
Aythya americana
31.0
25.6
27.3
25.8
31.5
25.3
27.6
25.8
19.1
24.5
22.5
24.2
18.4
24.6
22.5
24.2
37.5
49.1
45.0
48.4
James D. Watson & Francis H. Crick - 1953
Double Helix Model of DNA
Two sources of information:
2.
X-ray diffraction studies - Rosalind Franklin & Maurice Wilkins
Conclusion-DNA is a helical structure with
distinctive regularities, 0.34 nm & 3.4 nm.
Double Helix Model of DNA: Six main features
1.
Two polynucleotide chains wound in a right-handed (clockwise)
double-helix.
2.
Nucleotide chains are anti-parallel:
3.
Sugar-phosphate backbones are on the outside of the double
helix, and the bases are oriented towards the central axis.
4.
Complementary base pairs from opposite strands are bound
together by weak hydrogen bonds.
5’  3’
3’  5’
A pairs with T (2 H-bonds), and G pairs with C (3 H-bonds).
e.g.,
5’-TATTCCGA-3’
3’-ATAAGGCT-3’
5.
Base pairs are 0.34 nm apart. One complete turn of the helix
requires 3.4 nm (10 bases/turn).
6.
Sugar-phosphate backbones are not equally-spaced, resulting
in major and minor grooves.
1962: Nobel Prize in Physiology and Medicine
James D.
Watson
Francis H.
Crick
Maurice H. F.
Wilkins
What about?
Rosalind Franklin
RNA (A pairs with U and C pairs with G)
Examples:
mRNA
tRNA
rRNA
snRNA
messenger RNA
transfer RNA
ribosomal RNA
small nuclear RNA
RNA secondary structure:
Yeast Alanine tRNA
single-stranded
Function in
transcription
(RNA processing)
and translation
Organization of DNA/RNA in chromosomes
Genome = chromosome or set of chromosomes that contains all the
DNA an organism (or organelle) possesses
Prokaryotic chromosomes
1. most contain one double-stranded circular
DNA molecule
2. typically arranged in arranged in a dense
clump in a region called the nucleoid
Eukaryotic chromosomes
1. Eukaryotic chromosome structure
Chromatin - complex of DNA and chomosomal
proteins ~ twice as much or more protein as
DNA.
2. Eukaryotic chromosomes or chromatin found in the
nucleus of the cell.
3. Cells from different species contain varying numbers
of chromosome of different sizes
and morphologies -the karyotype (e.g., pea, 2N =
14; human, 2N = 46, fruit fly, 2N= 8).