DNA &

DNA REPLICATION
 Deoxyribonucleic Acid (DNA)

▪ Determines the genetics make-up of

living organisms
▪ Directs synthesis of proteins
▪ Enables cells to reproduce or make
other copies of themselves
 DNA Structure

▪Is made up of two strands to form a

double helix structure
▪Each strand is made up of repeating units
called nucleotides
▪Nucleotides consists of pentose sugar -
a five-carbon sugar, ribose or
deoxyribose, nitrogenous base and
phosphate
 DNA Structure
• Nitrogenous bases – Adenine (A),
Guanine (G), Cytosine (C), Thymine (T).
▪ A to T, by two hydrogen bonds
▪ C to G, by three hydrogen bonds
▪ Nucleotides are joined together by
phosphodiester bond
▪ 2 nucleotides – dinucleotides
▪ Many - polynucleotides
Nucleotide
 Base pairs

Non-covalent hydrogen bonds between

the pairs are shown as dashed lines.
DNA – Antiparallel strands
 DNA Structure
▪Higher GC content results in higher
melting temperatures
▪the genomes of extremophile organisms
such as Thermus thermophilus are
particularly GC-rich.
▪On the other hand, regions of a genome
that need to separate frequently — for
example, the promoter regions for
often-transcribed genes — are
comparatively GC-poor.
 Ribonucleic Acid (RNA)
• Ribonucleic acid (RNA) is a linear
molecule composed of four types of smaller
molecules called ribonucleotide bases:
adenine (A), cytosine (C), guanine (G), and
uracil (U).
• A to U, C to G.
• Ribose sugar
• A few types ; mRNA, tRNA, rRNA
 Messenger RNA (mRNA)
▪ A single-stranded molecule that is
transcribed from a DNA coding strand.
▪ Originates in the nucleus and migrate to
the ribosome in the cytoplasm in
preparation for the process protein
synthesis
▪ Consists of a sequence of mRNA
nucleotide (several thousands)
 Transfer RNA (tRNA)

▪Transcribed from DNA

▪A single strand, only about 80
nucleotides long and has a cloverleaf
shape
▪Function – to carry amino acids that will
be linked into the polypeptide molecule
specified by a particular sequence of
bases on the mRNA
 Transfer RNA (tRNA)
• Each type of tRNA carries only one of the
20 amino acids.
• Some can be carried by more than one
tRNA molecule
• The other end of tRNA molecule has three
nucleotides (the anticodon) which attach to
the matching set of three nucleotides
(codon) on the mRNA molecule.
Transfer RNA (tRNA)
 Ribosomal RNA (rRNA)

• ribosomal ribonucleic acid (rRNA) is

the RNA component of the ribosome, and
is essential for protein synthesis in all
living organisms. It constitutes the
predominant material within the ribosome,
which is approximately 60% rRNA and
40% protein by weight.
Ribosomal RNA (rRNA)
Central Dogma

Fig. 10-1, p.241

 DNA replication
•Purposes - to ensure that each cell in an
organism has a complete and correct
copy of the organism's genome.
•Occurs prior to cell division.
•Three important challenges ; separating
the two DNA strands, synthesizing of
DNA from 5’ to 3’ end guarding against
errors in replication (ensuring the correct
base is added to the growing
polypeptide chain)
 Semi-conservative Replication

•Proposed by Matthew Meselson and

Franklin Stahl in the late 1950s.
•Grown E. coli with 15NH4Cl (usual isotope of
N is 14N).
•All newly formed nitrogen compounds,
including purine and pyrimidine
nucleobases, become labeled with 15N.
•The 15N-labeled cells were then transferred
to a medium that containing 14N only.
 Semi-conservative Replication
•Analyzed DNA of E. coli (for every new
generation) using density-gradient centrifugation
technique.
•15N is a heavy isotope than that of 14N
•15N DNA forms a band at the bottom of the
tube, 14N DNA at the top of the tube, 15DNA and
14
DNA appears at a position halfway between
the two bands
•50-50 hybrid DNA was observed after one
generation, suggesting that semiconservative
replication had occurred.
https://www.
youtube.com
/watch?v=Je
oegQaF8ig

Fig. 10-2, p.242

 DNA replication in Prokaryotes
•Unicellular organism, example bacteria, E.
coli
•Circular DNA
•Prokaryotic DNA exists in supercoiled
(closed-circular form: tertiary structure of
DNA).
•two replication forks can proceed
independently around the DNA ring and when
viewed from above it resembles the Greek
letter "theta" (θ)
• Replication in
prokaryotes
• Two replication fork
• Theta replication

Fig. 10-4a, p.244

 Replication forks & bidirectional
replication
• Replication begins at specific site on
bacterial chromosome (origin)
• It starts at specific site (oriC) on bacterial
chromosome
• Proteins bind at oriC to initiate replication;
like promoter for RNA polymerase
• Replication moves out from origin in both
directions (bidirectional); forks meet across
from origin, where replication is terminated
• New duplexes separate
 DNA replication in eukaryotes
•Even though many of the principles are the
same, eukaryotic replication is more
complicated in three ways;
multiple origins of replication,
the timing must be controlled to that of cell
division
involved many proteins and enzymes
 DNA replication in eukaryotes
•Two replication fork on each origin
•The “bubbles that arises from each origin
eventually coalesce (combine and form
one group)
 DNA replication in Eukaryotes
Polymerases building both strands move in 3' to 5'
direction along template
One DNA strand grows toward replication fork;
the other away from fork; both grow 5'—>3'
Strand growing toward fork grows continuously in
5'—>3' direction (leading strand)
Strand growing away from fork grows
discontinuously as fragments (lagging strand);
initiation of each fragment must wait for parental
strands to separate & expose additional template
Some DNA was made as small segments
(Okazaki fragments) & rapidly joined to longer
DNA synthesized earlier by DNA ligase.
Fig. 10-5a, p.245
Fig. 10-5b, p.245
https://www.youtube.com/watch?v=Cw8GAPuhAk4

Fig. 10-10, p.249

Possible types of replication:

• Semiconservative
• Conservative
• Dispersive
 Semiconservative
• daughter duplex made of one parental & one newly
synthesized strand
 Conservative
• 2 original strands stay together after
serving as templates for 2 new strands
that also stay together; one contains only
"old" DNA, the other only "new" DNA
 Dispersive

• parental strand integrity disrupted;

new duplex strands made of old &
new DNA; neither the parental
strands nor the parental duplex is
preserved
Three models of DNA replication
Table 10-3, p.250
Table 10-5, p.260
END OF LECTURE