Professional Documents
Culture Documents
of Organisms: Part IV
DNA
Nucleic acids store and transmit hereditary information
3 C
Nucleoside
Nitrogenous
base Cytosine (C) Thymine (T, in DNA) Uracil (U, in RNA)
Purines
Phosphate
group Sugar
5 C (pentose)
Adenine (A) Guanine (G)
3 C (b) Nucleotide
Sugars
3 end
(a) Polynucleotide, or nucleic acid
5'C
3'C
Nucleoside
Nitrogenous
base
5'C
Phosphate 3'C
group Sugar
5'C (pentose)
3' end
(a) Polynucleotide, or nucleic acid
Nitrogenous bases
Pyrimidines
Purines
Sugar-phosphate
backbones
Old strands
Nucleotide
about to be
added to a
new strand
3'end
5'end
New
strands
3'end 5'end
5'end 3'end
!
"#
Secondary Structure
The secondary structure of DNA was proposed by
Watson and Crick in 1953
E. Chargaff noted that in DNA the percentage of
pyrimidine bases was approximately equal to the
percentage of purine bases
Also the mole percentage of adenine Is nearly
equal to that of thymine
The mole percentage of guanine is nearly equal to
cytosine
Chargaff also noted that the ratio of A and T versus G
and C varies by species but the ratio is the same for
different tissues in the same organism
X-ray crystallographic data showed the bond lengths
and angles of purine and pyrimidine bases
X-ray data also showed DNA had a long repeat distance (34
Å)
Based on this data, Watson and Crick proposed the
double helix model of DNA (next slide)
Two nucleic acid chains are held together by hydrogen
bonding between the bases on opposite strands
The double chain is wound into a helix
Each turn in the helix is 34Å long and involves 10 successive
nucleotide pairs
Each base pair must involve a purine and a pyrimidine to
achieve the proper distance between the sugar-phosphate
backbones
Base pairing can occur only between thymine and adenine, or
cytosine and guanine; no other pairing has the optimum
pattern of hydrogen bonding or would allow the distance
between sugar-phosphate backbones to be regular
X-ray diffraction photograph of a DNA fibre
(Rosalind Franklin and Maurice Wilkins)
Isolated nucleotides > single stranded DNA or RNA > double stranded DNA
Increasing supercoiling
Flow of Genetic Information
Transcription Translation
RNA and Protein Synthesis
“The central dogma of molecular genetics”
1 Synthesis of
mRNA in the
nucleus mRNA
NUCLEUS
CYTOPLASM
DNA
1 Synthesis of
mRNA in the
nucleus mRNA
NUCLEUS
CYTOPLASM
mRNA
2 Movement of
mRNA into cytoplasm
via nuclear pore
DNA
1 Synthesis of
mRNA in the
nucleus mRNA
NUCLEUS
CYTOPLASM
mRNA
2 Movement of
mRNA into cytoplasm Ribosome
via nuclear pore
3 Synthesis
of protein
Amino
Polypeptide acids
Transcription: Synthesis of Messenger RNA
(mRNA)
In the nucleus a DNA molecule partially unwinds
to expose a portion corresponding to at least one
gene
Ribonucleotides with complementary bases
assemble along the DNA strand
Base-pairing is the same in RNA, except that
in RNA uracil replaces thymine
Ribonucleotides are joined into a chain of mRNA
by the enzyme RNA polymerase
An intron (intervening sequence) is a segment of DNA
which is transcribed into mRNA but not actually used
when a protein is expressed
An exon (expressed sequence) in the part of the DNA
gene which is expressed
Each gene usually contains a number of introns and
exons
Introns are excised from mRNA after transcription
Ribosomes - rRNA
Protein synthesis is catalyzed in the cytoplasm by
ribosomes
–A ribosome consists of approximately two thirds
RNA and one third protein
–A ribosome is a ribozyme ( an reaction catalyst
made of ribonucleic acid)
A ribosome has 2 large subunits
–The 30S subunit binds the mRNA that codes for
the protein to be translated
–The 50S subunit catalyzes formation of the amide
bond in protein synthesis
Transfer of an amino acid to the growing peptide
chain is aided by acid-base catalysis involving an
adenine in the 50S subunits
Transfer RNA (tRNA)
Transfer RNAs (tRNAs), specific to each amino acid,
transport amino acids to complimentary binding sites on
the mRNA bound to the ribosome
–More than one tRNA codes for each amino acid
tRNA is comprised of a relatively small number of
nucleotides whose chain is folded into a structure with
several loops
–One arm of the tRNA always terminates in the
sequence cytosine-cytosine-adenine, and it is here the
amino acid is attached
–On another arm is a sequence of three bases called
the anticodon, which binds with the complementary
codon on mRNA
The mRNA genetic code is shown on the next slide
The Genetic Code
The genetic code is based on three-base sequences in
mRNA
Each three-base sequence corresponds to a particular
amino acid
–The fact that three bases are used to code for each
amino acid provides redundancy in the overall code
and in the start and stop signals
–N-formyl methionine (fMet) is the first amino acid
incorporated into bacterial protein and appears to be
the start signal
–fMet is removed from the protein chain before its
synthesis is complete
Translation
Translation is peptide synthesis by a ribosome
using the code from an mRNA
The polypeptide begins to acquire its secondary
and tertiary structure as it is being synthesized
Several ribosomes can be translating the same
mRNA molecule simultaneously
Protein molecules are synthesized only when they
are needed
–Regulator molecules determine when and if a
particular protein will be expressed i.e.
synthesized
Amplifying DNA in Vitro: The
Polymerase Chain Reaction (PCR)
The polymerase chain reaction, PCR, can
produce many copies of a specific target
segment of DNA
A three-step cycle—heating, cooling, and
replication—brings about a chain reaction
that produces an exponentially growing
population of identical DNA molecules
$ % &'() 5′′ 3′′
TECHNIQUE
Target
sequence
3′′ 5′′
2 Annealing
Cycle 1
yields Primers
2
molecules
3 Extension
New
nucleo-
tides
Cycle 2
yields
4
molecules
Cycle 3
yields 8
molecules;
2 molecules
(in white
boxes)
match target
sequence
$ % &'()
5′′ 3′′
TECHNIQUE
Target
sequence
3′′ 5′′
2 Annealing
Cycle 1
yields Primers
2
molecules
3 Extension
New
nucleo-
tides
$ % &'()
Cycle 2
yields
4
molecules
$ % &'()
Cycle 3
yields 8
molecules;
2 molecules
(in white
boxes)
match target
sequence
Gel Electrophoresis
One indirect method of rapidly analyzing and
comparing genomes is gel electrophoresis
This technique uses a gel as a molecular
sieve to separate nucleic acids or proteins by
size
A current is applied that causes charged
molecules to move through the gel
Molecules are sorted into “bands” by their size
$ % &'(*
TECHNIQUE
Mixture of Power
DNA mol- source
ecules of – Cathode Anode +
different
sizes
Gel
1
Power
source
– +
Longer
molecules
2 Shorter
molecules
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RESULTS
DNA and Proteins as Tape
Measures of Evolution
The linear sequences of nucleotides in DNA
molecules are passed from parents to
offspring
Two closely related species are more similar
in DNA than are more distantly related
species
Molecular biology can be used to assess
evolutionary kinship
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