Nucleic Acids

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Nucleic Acids

Stoker Chapter 22
Lippincott Chapters 30, 31 and
32

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1
Reserved.
2 Introduction

– Cells in an organism are capable of replicating


– Cells possess information on how to make new
cells
– Molecules responsible for such information are
nucleic acids
– Found in nucleus and are acidic in nature
– A nucleic acid is an unbranched polymer in
which the monomer units are nucleotides
Both DNA and RNA are unbranched polymers.

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3
Types of Nucelic
Acids

– Deoxyribonucleic acid (DNA)


– Found within the cell nucleus
– Stores and transfers genetic information
– Passed from existing cells to new cells
during cell division
– Ribonucleic Acid (RNA)
– Occurs in all parts of a cell
– Primary function is the synthesis of
proteins

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4
Nucleic Acids

– They are polymers in which the repeating unit is a


nucleotide
– Components of a nucleotide
– Pentose sugar
– Phosphate group (PO43-)
– Heterocyclic base Base

Phosphate Sugar
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5 Pentose Sugars

– The structural difference between


ribose and pentose 2′ -
deoxyribose occurs at carbon 2′
– —OH group is present in ribose
– —H atom is present in 2′-
deoxyribose
– Ribose is present in RNA
– 2′-deoxyribose is present in DNA

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•6 Nitrogen-Containing
Heterocyclic Bases

– They are five in total


– Three pyrimidine derivatives -
Thymine (T), cytosine (C), and uracil
(U)
– Two purine derivatives - Adenine
(A) and guanine (G)
• Adenine, guanine, and cytosine
are found in both DNA and RNA
• Uracil is found only in RNA
• Thymine is found only in DNA

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7

Phosphate – The third component of a nucleotide


– Derived from phosphoric acid (H3PO4)
– Loses two hydrogen atoms, resulting in
formation of a hydrogen phosphate ion
(HPO42–)

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8 Nucleotide Formation

– Nucleotides are formed from a


sugar, a base, and a phosphate
– It is a two-step process

Nucleoside is a substance that


– The pentose sugar and
does not contain a phosphate nitrogenous base react to form
group. Sugar and base pa lang
yung meron. a nucleoside
– The nucleoside reacts with a
phosphate group to form a
nucleotide

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Nucleoside
Formation

– Nucleoside: A two-subunit molecule in which


a pentose sugar is bonded to a nitrogen-
containing heterocyclic base
– Characteristics
– The base is attached to C1′ position of the sugar
(β-configuration)
– It is a condensation reaction

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9
10 Nucleoside Formation

– There are 8 nucleosides associated with nucleic acid chemistry


– Four RNA nucleosides
– Four DNA nucleosides
– Nomenclature
– For pyrimidine bases, the suffix -idine is used
– Cytidine, thymidine,
– For purine bases, the suffix -osine is used
– Adenosine, guanosine
– Prefix deoxy- is used to indicate deoxyribose present
– Deoxythymidine

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Nucleotide
Formation

– Formed by the addition of a phosphate group


to a nucleoside
– Characteristics of phosphate addition
– The phosphate is attached to C5′ position
– Water is released Dehydration

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11
Table 22.1 - The Building Blocks of DNA and
RNA

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12 Reserved.
13

Primary – A ribonucleic acid (RNA) is a


nucleotide polymer in which each of
Nucleic Acid the monomers contains ribose, a
Structure phosphate group, and one of the
heterocyclic bases adenine, cytosine,
guanine, or uracil
– A deoxyribonucleic acid (DNA) is a
nucleotide polymer in which each of
the monomers contains deoxyribose, a
phosphate group, and one of the
heterocyclic bases adenine, cytosine,
guanine, or thymine

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Primary
Nucleic Acid
Structure
– A sequence that
contains nucleotides
linked to a nucleic acid
– Primary structure
depends on the
sequence of bases
present

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14
Figure 22.6 - Comparison of the General Primary
Structures of Nucleic Acids and Proteins

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15 Reserved.
16 The DNA Double Helix

– In addition to primary structures, nucleic acids also have secondary


and tertiary structures
– Amounts of A,T,G, and C present in DNA molecules helped determine
the three-dimensional structure of the DNA molecules
– Amounts of A and T were always equal
In a normal DNA double helix, the adenine and thymine is always equal.
– Amounts of C and G were always equal
– Human DNA contains:
– 30% adenine
– 30% thymine
– 20% guanine
– 20% cytosine

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Figure
22.7 -
The DNA
Double
Helix

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Reserved.
18 DNA Sequence

– Two polynucleotide strands are coiled around


each other in a spiral
– Bases on each strand extend inwardly toward each
other
– They are connected by hydrogen bonds
– They run in opposite directions Also called anti-parallelism

– 5′-to-3′ direction
– 3′-to-5′ direction

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19
Base Pairing

– A pyrimidine is always paired with a purine


– Fits inside the DNA double strand
– Hydrogen bonding is most favored in A–T
and G–C pairs
– A–T and G–C pairing is termed
complementary

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20 Practice Exercise

– Predict the sequence of bases in


the DNA strand complementary to
the single DNA strand shown
below:

5′ A–A–T–G–C–A–G–C–T 3′

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21
Practice Exercise

– Predict the sequence of bases in the DNA strand


complementary to the single DNA strand shown below:

5′ A–A–T–G–C–A–G–C–T 3′

Answer:
3′ T–T–A–C–G–T–C–G–A 5′

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22 Base Pairing

– Hydrogen Bonding Interactions Weak

– Base-Stacking Interactions (like


stack of coins) Makes it look like they’re strong

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23 Replication of DNA Molecules

– DNA replication: The biochemical process by which DNA molecules


produce exact duplicates of themselves
– The strands of DNA are used as templates
– DNA polymerase ensures correct base pairing and catalyzes the
formation of phosphodiester linkages
– Components of a new DNA strand
– One new DNA strand
– Daughter strand
– One old DNA strand
– Parent strand

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24 The Replication Process in Finer
Detail

– DNA polymerase enzyme can operate on a forming DNA daughter strand only in
the 5′-to-3′ direction
– One strand grows continuously in the 5′-to-3′ direction
– The other strand grows in segments in the opposite direction
– Okazaki fragments
– The segments are connected by DNA ligase
– DNA replication usually occurs at multiple sites within the molecule
– Bidirectional replication occurs at these sites
– Multiple-site replication enables rapid replication of large molecules

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Figure 22.1 - Continuous-Growing Strands and
Okazaki Fragments

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25 Reserved.
26 Chromosomes

– They are formed by the interaction of newly-replicated


DNA molecules with histones
– A chromosome is about 15% by mass DNA and 85% by
mass protein
– Cells of different kinds of organisms have different
numbers of chromosomes
– They occur in matched (homologous) pairs
‒ Example - The 46 chromosomes of a human cell constitute 23
homologous pairs

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27
Overview of Protein Synthesis

– DNA molecules control the synthesis of proteins


– Phases of protein synthesis
– Transcription
– Translation

Transcription Translation
DNA RNA Protein

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28
Differences Between RNA
and DNA Molecules
– DNA Molecule – RNA Molecule

– Deoxyribose is the basic – Ribose is the basic sugar


sugar unit unit

– Thymine is present – Thymine is replaced by


uracil
– It is a double-stranded
helix – It is a single-stranded unit

– Molecules are bigger – Molecules are smaller

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29 Types of RNA Molecules

– Heterogenous nuclear RNA (hnRNA): Formed by DNA


transcription
– Messenger RNA (mRNA): Carries instructions for protein
synthesis
– Small nuclear RNA (snRNA): Facilitates the conversion of
hnRNA to mRNA
– Ribosomal RNA (rRNA): Combines with specific proteins to
form ribosomes

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Types of RNA Molecules
Transfer RNA (tRNA): Delivers amino acids to sites for protein synthesis

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30 Reserved.
•31 Transcription

– Transcription: The process by which DNA directs the synthesis of


hnRNA/mRNA molecules by a two-step process
– hnRNA is synthesized
– hnRNA is edited to yield mRNA
– Gene: A segment of a DNA strand responsible for the production
of a specific hnRNA/mRNA molecule
– Most human genes comprise 1000–3500 nucleotide units
– Genome: All of the genetic material contained in the
chromosomes of an organism

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32 Steps in the Transcription
Process

– Unwinding of DNA double helix to expose a sequence


of bases
– Governed by RNA polymerase
– Alignment of free ribonucleotides along an exposed
DNA strand, forming new base pairs
– Influence of RNA polymerase in the linkage of
ribonucleotides
– Cessation of transcription when the RNA polymerase
enzyme encounters a stop signal on the DNA
template
– The newly formed RNA molecule and the RNA
polymerase enzyme are released

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33 Steps in the
Transcription Process

– The strand of DNA used for hnRNA/mRNA


synthesis is called the template strand. It is
copied proceeding in the 3’ to 5’ direction.
– The other DNA strand (the non-template strand)
is called the informational strand.
– The informational strand, although not involved
in RNA synthesis, gives the base sequence
present in the hnRNA strand being synthesized
(with the exception of U replacing T).

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34
Post-Transcription
Processing: Formation of
mRNA
– Involves post-transcription processing of hnRNA
– Splicing: Excision of introns and joining of exons
– Exon: A gene segment that conveys codes for
genetic information
– Intron: A DNA segment that does not convey
genetic information
– snRNA is involved

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Alternative
splicing

– Alternative splicing: A
process of producing
several different
proteins from a single
gene
– Involves splicing of
an hnRNA molecule
with multiple exons

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35
36 Transcriptome

– All of the mRNA molecules that can be


generated from the genetic material in a genome
– It is different from a genome
– Acknowledges the biochemical complexity
created by splice variants obtained by hnRNA

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The Genetic Code
37

– The base sequence in mRNA determines the amino acid


sequence in the synthesis of protein
– The base sequence of an mRNA molecule involves 4
different bases - A, C, G, and U
– Codon: A three-nucleotide sequence in an mRNA
molecule that codes for a specific amino acid
– 64 codons in total
– Genetic code: The assignment of the 64 mRNA codons
to specific amino acids
– 3 of the 64 codons are termination codons

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•3 Characteristics of
8
Genetic Code

– The genetic code is highly degenerate


– Many amino acids are designated by more
than one codon
– Met and Trp possess a single codon
– There is a pattern to the arrangement of
synonyms in the genetic code table
– All synonyms for an amino acid fall within
a single category unless there are more
than four synonyms

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39
Characteristics of
Genetic Code

– The genetic code is almost universal


– There are minor exceptions
– An initiation codon exists
– Suggested by the existence of “stop”
codons
– AUG is an initiator of protein synthesis

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Anticodons and tRNA Molecules

– During protein synthesis, amino acids do not directly interact with the codons of an
mRNA molecule
– tRNA molecules act as intermediaries to deliver amino acids to mRNA
– Two important features of the tRNA structure
– The 3′ end of tRNA is where an amino acid is covalently bonded to the tRNA
– The loop opposite to the open end of tRNA, called the anticodon, comprises seven
unpaired bases
– Three unpaired bases constitute the anticodon
– A three-nucleotide sequence on a tRNA molecule that is complementary to a
codon on an mRNA molecule

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Translation: Protein Synthesis

– Translation: The process in which mRNA codons are deciphered and a specific protein molecule is
synthesized
– Ribosome: An rRNA–protein complex that is the site for the translation phase of protein synthesis
– Characteristics of ribosome structures
• They contain four rRNA molecules and 80 proteins in two subunits
• Each subunit possesses 65% rRNA and 35% protein
• The active site is located in the ribosomal subunit
• rRNA is the active site
• The predominance of rRNA at the active site gives it the impression of a ribozyme
• The mRNA binds to the small subunit of the ribosome

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Five Steps of Translation

– Activation of tRNA
– Accomplished in two steps
– Initiation
– tRNA attaches itself to the P site of a small ribosomal unit
– Elongation
– Another tRNA attaches itself to the A site
– A dipeptide is formed under the influence of peptidyl transferase
– Termination
– The polypeptide continues to grow via translocation till a stop codon is encountered
– Post-translational processing
– The protein is rendered fully functional

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43 Efficiency of mRNA
Utilization

– Many ribosomes can move


simultaneously along a single
mRNA molecule
– The multiple use of mRNA
molecules reduces the amount of
resources and energy that the cell
expends to synthesize needed
protein
– Polyribosome: A complex of an
mRNA and several ribosomes

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44 Mutation

– An error in base sequence in a gene that is


reproduced during DNA replication
– Altered genetic information is passed on during
transcription
– Altered information can cause changes in amino
acid sequence during protein synthesis
– Changes can cause a profound effect on the
organism

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A mutagen is a substance or
agent that causes a change in the
structure of a gene
45

Mutagens
Radiation

Types •Ultraviolet light, X-


rays, and radioactivity
Chemical agents

HNO2

Repair enzymes present in the


body are able to identify and
replace altered bases
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Viruses
Viruses are categorized depending on the type of nucleic acid present

– Minute disease-causing agents with an outer coat of


protein
– They can reproduce only by invading host cells
Which means outside the body, viruses are actually dead.
– Host cells are caused to produce more viruses
– Host cells’ normal function is disrupted
– They attack bacteria, plants, animals, and humans
– Many human diseases are of viral origin

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46
47
– They attach themselves to the host cell on
the external surface
– An enzyme present in the external structure
of the virus catalyzes the breakdown of the
cell membrane and forms a hole
Viruses – They then inject their DNA or RNA into the
host cell
– The viral nucleic acid is replicated
– Hundreds of new viruses are produced
using the host cell within 30 minutes

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48 Vaccine

– A preparation containing an inactive or weakened form of a


virus
– Antibodies produced against inactive viral or bacterial
envelopes also kill naturally occurring viruses or bacteria

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•49 Recombinant DNA and Genetic
Engineering

– Genetic engineering: The process by which an organism is


intentionally changed at the molecular (DNA) level so that
it exhibits different traits
– Recombinant DNA: DNA possessing genetic material from
two different organisms

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50 Recombinant DNA and Genetic
Engineering

– First genetically engineered organisms were


bacteria (1973) and mice (1974)
– Insulin-producing bacteria were
commercialized in 1982
– Many plants have now been genetically
engineered
– Resistance to the harmful effects of
pesticides
– Protection from insects
– Plants produce pesticides
– Increased crop production

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Principles and Procedures of Genetic
Engineering

– E.coli cells containing recombinant DNA are obtained using the


following steps
– Cell membrane dissolution
– Isolation of plasmid fraction
– Cleavage of plasmid DNA
– Gene removal from another organism
– Gene-plasmid splicing
– Uptake of recombinant DNA

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52 Principles and Procedures of
Genetic Engineering

– Transformed cell can reproduce a large


number of identical cells called clones
– Cells with identical DNA that have
descended from a single cell
– Thousands of clones can be produced
from a single bacteria within a short
time
– Each clone can synthesize the protein
directed by foreign gene it carries

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Figure 22.30
- Production
of
Recombinant
DNA

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53 Reserved.
A method for rapidly
producing multiple copies of 54
a DNA nucleotide sequence
The
Polymerase
Chain
Reaction
This method allows to
produce billions of copies of a
DNA sequence in a few hours

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Figure 22.33 (a) - Basic Steps of the Polymerase
55 Chain Reaction

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Figure 22.33 (b) - Basic Steps of the Polymerase Chain
56

Reaction

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