DNA and the processing of RNA

The molecular basis of Heredity

• James Watson & Francis Crick (April 1953)
publish the double-helix model for DNA
• Two fundamental genetic mechanisms:
1. Storage of genetic information in a linear,
deadagle pattern
2. Replication of genetic information to ensure its
faithful transmission from generation to
generation.
A. DNA as carrier of genetic information
1. Investigation of evidence of DNA

• Fredrick Griffith (1928) while investigating several

strains of the bacteria Streptococcus pneumoniae he
observed the phenomena “transformation”
• Oswald Avery et al. (1944) determined that the chemical
basis of the transforming principle is DNA
• The results could not be explained - sceptism (its
relevance for genetics was not apparent)
Transformation of bacteria

Fredrick Griffith (1928) described by the bacteria

Streptococcus pneumoniae the phenomenia “transformation”
by a “foreign substantion”
A. DNA: genetic material
2. Research to prove the existance of DNA

• Studies with bacteriophages: bacteria infected by virusses

• Hershey & Chase (1952) demonstrated that DNA was the
genetic material of the phage T2 that infected Escherichia
coli
• He investigated this by his experiment with 35S (only in
proteins) en 32P (only in DNA)
The Hershey-Chase experiment: phages on the surface of a bacterial cell
The Hershey-Chase experiment
A. DNA as genetisch materiaal
2. Investigation of evidence of DNA

• Evidence for DNA as genetic material: observations in

eukaryotes:
• DNA multiplication before mitosis
• Diploid cells contain 2x more DNA than the gametes
• Chargaff (1947) developed the basic rules for A, C, G, T
• % Thymine ~= % Adenine
• % Guanine ~= % Cytosine
Structure of a single DNA strand
Rosalind Franklin: the X-ray diffraction photo of DNA
Base paring in DNA by Watson & Crick (respecting the Chargaff basic rule)
De double helix : 3 ways of presenting
 B. DNA-replication
Basepairs serve as a template
• Watson & Crick published their basic concept
about DNA replication
• Single stainded DNA serve as a template for a
new complementary strand.
B. Mechanism: DNA-replication

• Fast – ‘relatively’ few errors

• E. coli : ca. 500 -1000 bp per second
• Human: ca. 50 bp per second

• Start of replication by “bubble”-forming

in bacteria: 1 site
in eucaryotes: >1000 sites
Origin of replication in eukaryotes: replication-enzymes separate the strains
The two strands of the DNA molecule are antiparallel
Building a new strand
A model for DNA replication: the basic concept
C. DNA-replication and DNA-repair:
an overview:
• Leading strand 3’-> 5’
• Lagging strand 5’-> 3’ with Okazaki fragments
• Okazaki fragments (100-200 nucleotiden) fixed with
DNA ligase
• DNA-polymerase adds the nucleotiden
• The start requires a primer (short RNA fragment)
• Primase (RNA polymerase) attaches to complementary
ribonucleotides at the DNA template
• Afterwards DNA polymerasen attach to deoxyribonucleotides
The “leading strand” and the “lagging strand” during DNA replication

LEADING
STRAND

LAGGING STRAND
Starting up DNA synthesis with the RNA primer
Overview of DNA-replication: the replication fork
The most important proteins in the process of DNA replication and their functions
Bacterial DNA replication proteins and their functions
D. Mismatch & nucleotide excision repair
• Ca. 1 error per 10.000 bp by reactive chemicalien,
radioactive radiation, X-radiation, UV-radiation :
DNA polymerase stops replication
• Ca. 130 repair enzymesin human
• Mismatch repair (ex. T-T dimeren)
• Nucleotide excision repair (nucleases + DNA-
polymerase + ligase)
“Nucleotide excision repair “ of DNA damage
Replication at the ends of DNA strands
E. Telomeres at the ends of DNA strains

• At each cell division somatic cells lose

nucleotides from their DNA strains. Problems at
the end of 5’ ends of daughter strains DNA
polymerase is limited
• The ends of chromosomen own special nucleotide
sequenties : telomeres
• Vb. TTAGGG (100-1000 repeats bij mens)
Telomeres, the aging clock in every cell
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Telomeres and telomerase: Telomeres on the chromosoms of the mouse

Telomereres are repeated along about 60kb and are present in the
germ cells (photo: zygote with large telomers) and in cancer cells (they
divide continuously)
No telomeric activity is found in somatic cells (tissues)