NUCLEIC ACIDS

An INTRODUCTION

Two classes of Nucleic Acids

Deoxynucleic Acids (DNA)

Hereditary molecule of all cellular life
Stores genetic information (encodes)
Transmits genetic information
Information on protein sequence is encoded
in DNA and translated by the help of
messenger Ribonucleic Acid (mRNA)
Ribonucleic Acids
Encode and Translate information on DNA to
proteins

1
 Composition and Structure of
Nucleic Acids
Nucleobases
Derivatives of purine and pyrimidine bases
Sugars
Phosphoric acid
DNA nucleobases
Adenine (A) and Guanine (G)
Tymine (T) and Cytosine (C)
RNA nucleobases
A and G
Uracyl (U) and Cytosine (C)

Nucleobases
6
7
1 5
2

3 9 1

Uracyl thymine without the methyl group

DNA nucleobases
Adenine (A) and Guanine (G)
Tymine (T) and Cytosine (C)

RNA nucleobases
A and G
Uracyl (U) and Cytosine

2
 Sugars
-D-ribose and -D-deoxyribose

DNA Nucleosides

Base (N9 or N1 atom) + sugar (C1atom)

N-glycosidic bond

3
 Building Blocks: Nucleotides

Nucleoside +
phosphate
group(s)
5-hydroxylic
group of sugar
phosphate
Ester bond

Polynucleotides
A T G C
sugar phosphate sugar phosphate sugar phosphate sugar ...

Backbone : sugar + phosphate

Side groups: nucleobases
Reading from
From Nucleotide with free phosphate
group (5end)
To nucleotide with free 3hydroxyl group
(the 3 end)

4
 Double Strand Formation
Base Pairing

Double strand DNA is inherently very sensitive/ fragile

Strands Pairing

5
 The Double Helix

Denatured during replication and mRNA synthesis

Denatured during PCR (amplification of DNA)

Levels of Structural Organization

Primary
Sequence of nucleotide
Name of nucleobases used : ATCT
Secondary
Double stranded helix structure
Tertiary
Folding of the double helix
Example: super coiled

6
 Structure of RNA
Single strand
Folds on itself thus allowing pairing of
bases

Synthesis of Proteins

Procaryote cell Eucaryote cell

7
 Synthesis of Proteins

Synthesis of Proteins
1. Transcription
One strand of the DNA double helix is used as a template by the RNA
polymerase to synthesize a messenger RNA (mRNA).
mRNA migrates from the nucleus to the cytoplasm.
mRNA goes through different types of maturation including one called
splicing when the non-coding sequences are eliminated.
The coding mRNA sequence can be described as a unit of three
nucleotides called a codon.
2. Translation
The ribosome binds to the mRNA at the start codon (AUG) that is
recognized only by the initiator tRNA.
The ribosome proceeds to the elongation phase of protein synthesis.
During this stage, complexes, composed of an amino acid linked to
tRNA, sequentially bind to the appropriate codon in mRNA by forming
complementary base pairs with the tRNA anticodon.
The ribosome moves from codon to codon along the mRNA. Amino
acids are added one by one, translated into polypeptidic sequences
dictated by DNA and represented by mRNA.
At the end, a release factor binds to the stop codon, terminating
translation and releasing the complete polypeptide from the ribosome.
One specific amino acid can correspond to more than one codon. The
genetic code is said to be degenerate.

8
 Synthesis of Proteins: Genetic Code

Protein Synthesis
Folding, disulfide bridges formation, post-
translational modification

Post-translational modifications
Not determined by DNA sequence
Cleavage of amino acids
Phosphorylation
Acetylation
glycosylation

9
 Analytical Challenges for the Analysis of Biological
Macromolecules

Inherent sensitivity

Both proteins and nucleic acid 3D structure are very

sensitive to environmental conditions such as pH,
ionic strength, solvent composition

Why? stabilized by H-bonds, other dipole-dipole

interactions, electrostatic interactions, van der Waals
interactions

Problems
Denaturation
Crystallization challenges

Analytical Challenges for the Analysis of Biological

Macromolecules
Size of macromolecules
Several signals that must be resolved and analyzed
Complex three dimensional Structure
NMR challenges

Number of macromolecules in real samples

Separation methods
Methods for selective detection

Amount of sample
Sensitivity must be high
Amplification

10