NUCLEIC ACID

HENNY RACHDIATI

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INTRODUCTION
• Friedrich Miescher in 1869
o isolated what he called nuclein from the nuclei of pus cells
o Nuclein was shown to have acidic properties, hence it became called
nucleic acid
• The most important clue to the structure of DNA came from
the work of Erwin Chargaff and his colleagues in the late 1940s.
o The data collected from DNAs of a great many different species led
Chargaff to some important conclusions Chargaff’s “rules”…

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INTRODUCTION
• Two types of nucleic acid are found
o Deoxyribonucleic acid (DNA)
o Ribonucleic acid (RNA)
• The distribution of nucleic acids in the eukaryotic cell
o DNA is found in the nucleus with small amounts in mitochondria and
chloroplasts
o RNA is found throughout the cell

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Chargaff’s “rules”…
• Erwin Chargaff Garrett Vaughn Christian Skinner (1905-2002)

1. in all cellular natural DNAs the number of guanine units

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Erwin Chargaff Data (1950-1951)

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Structure Nucleic Acid
• Nucleic acids are polynucleotides
• Their building blocks are nucleotides
• Two major classes of nucleic acids:
1. deoxyribonucleic acid (DNA): carrier of genetic information
2. ribonucleic acid (RNA): an intermediate in the expression of genetic
information and other diverse roles
• The Central Dogma (F. Crick):
DNA mRNA Protein
(genome) (transcriptome) (proteome)

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Structure Nucleic Acid

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Nucleotides and Nucleosides
• Nucleotides contain Base + sugar + phosphoryl group

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Nucleotides
• Nucleotides contain Base(N) + sugar + phosphoryl group
• Nucleotides = nucleoside + phosphate
• Result from linking one or more phosphates with a nucleoside
onto the 5’ end of the molecule through esterification

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Nucleotides

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Nucleotides
• Nucleotides contain Base(N) + sugar + phosphoryl group
• Nitrogenous Bases
Planar, aromatic, and heterocyclic
Derived from purine or pyrimidine
Numbering of bases is “unprimed

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Nucleotides
• Nucleotides contain Base(N) + sugar + phosphoryl group
• Sugars
Pentoses (5-C sugars)
Numbering of sugars is “primed”

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Nucleotides
• Nucleotides contain Base(N) + sugar + phosphoryl group
• Phosphoryl Groups
• Mono-, di- or triphosphates
• Phosphates can be bonded to either C3 or C5 atoms of the sugar

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Nucleotides
• RNA (ribonucleic acid) is a polymer of ribonucleotides
• DNA (deoxyribonucleic acid) is a polymer of deoxyribonucleotides
• Both deoxy- and ribonucleotides contain Adenine, Guanine and
Cytosine
o Ribonucleotides contain Uracil
o Deoxyribonucleotides contain Thymine
• Monomers for nucleic acid polymers
• Nucleoside Triphosphates are important energy carriers (ATP,
GTP)
• Important components of coenzymes
o FAD, NAD+ and Coenzyme A

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Nucleosides
• Nucleosides : result from linking one of the sugars with a purine
or pyrimidine base through an N-glycosidic linkage
• Purines bond to the C1’ carbon of the sugar at their N9 atoms
• Pyrimidines bond to the C1’ carbon of the sugar at their N1 atoms

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 Nucleosides
• Nucleosides : = carbohydrate + base
• To differentiate the atoms of the carbohydrate from the base, the position number of the
carbohydrate is followed by a ´ (prime).
• The stereochemistry of the glycosidic bond found in nucleic acids is β.

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 Nucleoside, nucleotides and nucleic acids
The chemical linkage between monomer units in nucleic acids is
a phosphodiester

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Naming
• Nucleosides:
• Purine nucleosides end in “-sine”
• Adenosine, Guanosine
• Pyrimidine nucleosides end in “-dine”
• Thymidine, Cytidine, Uridine
• Nucleotides:
• Start with the nucleoside name from above and add “mono-”,
“di-”, or “triphosphate”
• Adenosine Monophosphate, Cytidine Triphosphate, Deoxythymidine
Diphosphate

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Polynucleic acid
• Polynucleic acid chain =
connected nucleotides
binding by 3’5’phosphor
diester bonds
• 3’,5’phosphodiester
bridges link nucleotides
together to form
polynucleotide chains

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Properties of Nucleic Acid
1. General physical and chemical properties
a. Amphiphilic molecules; normally acidic because of phosphate.
b. Solid DNA white fiber; RNA white powder. Insoluble in organic
solvents, can be precipitate by ethanol.
c. Can be hydrolyzed by acid/alkaline/enzymes
2. UV absorption
a. Specific absorption at 260nm.
b. This can be used to identify nucleic acid

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Properties of Nucleic Acid
3. Denaturation
Concept:
the course of hydrogen bonds broken, 3-D structure was destroyed,
the double helix changed into single strand irregular coid
Results:
1. the value of 260 nm absorption is increased
2. Viscous is decreased
3. biological functions are lost

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Properties of Nucleic Acid
Heat denaturation and Temperature
When DNA were heated to certain temperature, the absorption value at
260nm would increased sharply which indicates that the double strand
helix DNA was separated into single strand. When the absorption value
increases to 40%, the value change would low down, which indicates the
double strands had been completely separated.

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Properties of Nucleic Acid
Heat denaturation and Temperature

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Properties of Nucleic Acid
Heat denaturation and Temperature
Tm: melting temperature of DNA
The temperature of UV absorption increase to an half of
maximum value in DNA denaturation.
Factors affect Tm:
G-C content: there are three hydrogen bonds between G-C
pair. The more G-C content, the higher Tm value.
 (G+C)% = (Tm-69.3) × 2.44

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Properties of Nucleic Acid

4. Renaturation of DNA

When slowly cooling down the denatured DNA solution, the

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Properties of Nucleic Acid

5. Molecule hybridization
During the course of lowing down denaturing temperature,
between different resource DNAs or single stand DNA and
mRNA with complementary bases will repair into a double
strands to form a hybrid DNA or DNA-RNA . This course is
called molecule hybridization.

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Properties of Nucleic Acid
5. Molecule hybridization

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NUCLEIC ACID
HENNY RACHDIATI

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