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Chapter 3 Structures and Functions of Nucleic Acids. Nucleic acid. A biopolymer composed of nucleotides linked in a linear sequential order through 3’,5’ phosphodiester bonds. Classification of nucleic acid. Ribonucleic acid (RNA) is composed of ribonucleotides .

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Chapter 3 structures and functions of nucleic acids

Chapter 3Structures and Functions of Nucleic Acids

Nucleic acid
Nucleic acid

A biopolymer composed of nucleotides linked in a linear sequential order through 3’,5’ phosphodiester bonds

Classification of nucleic acid
Classification of nucleic acid

  • Ribonucleic acid (RNA) is composed of ribonucleotides.

    • in nucleiand cytoplasm

    • participate in the gene expression

  • Deoxyribonucleic acid (DNA)is composed of deoxyribonucleotides.

    • 90% in nuclei and the rest in mitochondria

    • store and carry genetic information; determine the genotype of cells

Interesting history
Interesting history

  • 1944: proved DNA is genetic materials(Avery et al.)

  • 1953: discovered DNA double helix (Watson and Crick)

  • 1968: decoded the genetic codes (Nirenberg)

  • 1975: discovered reverse transcriptase (Temin and Baltimore)

  • 1981: invented DNA sequencing method (Gilbert and Sanger)

  • 1985: invented PCR technique(Mullis)

  • 1987: launched the human genome project

  • 1994: HGP in China

  • 2001: accomplished the draft map of human genome

Section 1 chemical components of nucleic acids

Section 1Chemical Components of Nucleic Acids

§ 1.1 Molecular Constituents

Nucleic acid can be hydrolyzed into nucleotides by nucleases. The hydrolyzed nucleic acid has equal quantity of base, pentose and phosphate.



nucleic acid




Adenine (A)

Guanine (G)

Base: Purine

Uracil (U)

Thymine (T)

Cytosine (C)

Base: Pyrimidine







glycosidic bond

Purine N-9 or pyrimidine N-1 is connected to pentose (or deoxypentose) C-1’ through a glycosidic bond.


phosphoester bond

A nucleoside (or deoxynucleoside) and a phosphoric acid are linked together through the 5’-phosphoester bond.

Nucleic acid derivatives




Nucleic acid derivatives

Multiple phosphate nucleotides

  • adenosine monophosphate (AMP)

    • adenosine diphosphate (ADP)

    • adenosine triphosphate (ATP)

Nucleic acid derivatives1


Nucleic acid derivatives

Cyclic ribonucleotide: 3’,5’-cAMP, 3’,5’-cGMP, used in signal transduction

Nucleic acid derivatives2
Nucleic acid derivatives

Biologically active systems containing ribonucleotide: NAD+, NADP+, CoA-SH

Phosphoester bond formation

The -P atom of the triphosphate group of a dNTP attacks the C-3’ OH group of a nucleotide or an existing DNA chain, and forms a 3’-phosphoester bond.

Nucleic acid chain extension

A nucleic acid chain, having a phosphate group at 5’ end and a -OH group at 3’ end, can only be extended from the 3’ end.

Phosphodiester bonds

Alternative phosphodiester bonds and pentoses constitute the 5’-3’ backbone of nucleic acids.

Section 2 structures and functions of nucleic acids

Section 2Structures and Functions of Nucleic Acids

2 1 primary structure
§ 2.1 Primary Structure

  • The primary structure of DNA and RNA is defined as the nucleotidesequence in the 5’ – 3’ direction.

  • Since the difference among nucleotides is the bases, the primary structure of DNA and RNA is actually the base sequence.

  • The nucleotide chain can be as long as thousands and even more, so that the base sequence variations create phenomenal genetic information.

2 2 secondary structure
§ 2.2 Secondary structure

The secondary structure is defined as the relative spatial position of all the atoms of nucleotide residues.

2 2 a chargaff s rules
§ 2.2.a Chargaff’s rules

  • The base composition of DNA generally varies from one species to another.

  • DNA isolated from different tissues of the same species have the same base composition.

  • The base composition of DNA in a given species does not change with its age, nutritional state, and environmental variations.

  • The molarity of A equals to that of T, and the molarity of G is equal to that of C.

Molarity of bases









E. coli






















































Molarity of bases

Building a milestone of life

James Watson and Francis Crick proposed a double helix model of DNA in 1953.

It symbolized the new era of modern biology.

2 2 b double helix of dna
§ 2.2.b Double helix of DNA

  • Two DNA strands coil together around the same axis to form a right-handed double helix (also called duplex).

  • The two strands run in opposite directions, i.e., antiparallel.

  • There are 10 base pairs or 3.4nm per turn and the diameter of the helix is 2.0nm.

Backbone and bases

The hydrophilic backbone is on the outside of the duplex, and the bases lie in the inner portion of the duplex.

Base interactions

  • The two strands of DNA are stabilized by the base interactions.

  • The bases on one strand are paired with the complementary bases on another strand through H-bonds, namely G≡C and A=T.

  • The paired bases are nearly planarand perpendicular to helical axis.

  • Two adjacent base pairs have base-stacking interactions to further enhance the stability of the duplex.

Groove binding

Small molecules like drugs bind in the minor groove, whereas particular protein motifs can interact with the major grooves.

2 2 c polymorphisms of dna
§ 2.2.c Polymorphisms of DNA

  • DNA can resume different forms depending upon their chemical microenvironment, such as ionic strength and relative humidity.

  • B-form DNA is the predominant structure in the aqueous environment of the cells.

  • A-form and Z-form are also native structures found in biological systems.

Hoogsteen base pair

The third strand is using Hoogsteen H-bonds to pair with bases on the first strand.

G-quartet DNA

  • The telomere of DNA is a G-righ sequence, such as

  • 5’ (TTGGGG)n 3’

  • 4 G residues constitute a plane which is stabilized by Hoogsteen H-bonds.

G-quartet of DNA

Four strands are arranged in either parallel or antiparallel manner.

2 3 supercoil structure
§ 2.3 Supercoil Structure

§2.3.a Supercoil structure

  • The two termini of a linear DNA could be joined to form a circular DNA.

  • The circular DNA is supercoiled, and supercoil can be either positive or negative.

  • Only the supercoiled DNA demonstrate biological activities.

Em image of supercoiled dna
EM image of supercoiled DNA

Circular DNAs in nature, in general, are negatively supercoiled.

§2.3.b Prokaryotic DNA

  • Most prokaryotic DNAs are supercoiled.

  • Different regions have different degrees of supercoiled structures.

  • About 200 nts will have a supercoil on average.

§2.3.c Eukaryotic DNA

  • DNA in eukaryotic cells ishighly packed.

  • DNA appears in a highly ordered form called chromosomes during metaphase, whereas shows a relatively loose form of chromatin in other phases.

  • The basic unit ofchromatin is nucleosome.

  • Nucleosomes are composed of DNA and histone proteins.


  • DNA: ~ 200 bps

  • Histone: basic proteins with many Lys and Arg residues

    • H2A (x2),

    • H2B (x2),

    • H3 (x2),

    • H4 (x2)

Beads on a string

  • 146 bp of negatively supercoiled DNA winds 1 ¾ turns around a histone octomer.

  • H1 histone binds to the DNA spacer.

The total length of 46 human chromosomes is about 1.7 m, and becomes 200 nm long after 5 times condensation.

2 4 functions of dna
§ 2.4 Functions of DNA

DNA is fundamental to individual life in terms of

  • They are the material basis of life inheritance, providing the template for RNA synthesis.

  • They are the information basis for biological actions, carrying the genetic information.

  • DNA is able to replicate itself in a high fidelity to ensure the genetic information transfer from one generation to the next.

  • DNA can be used as a template to synthesize RNA (transcription), and RNA is further used as the template to synthesize proteins (translation).

  • DNA posses the inherent and the mutant properties to create the diversity and the uniformity of the biological world.

Gene and genome
Gene and genome

  • A gene is defined as a DNA segment that encodes the genetic information required to produce functional biological products.

  • A gene includes coding regions as well as non-coding regions.

  • Genome is a complete set of genes of a given species.

Section 3 structures and functions of rna

Section 3Structures and Functions of RNA


  • mRNA (messenger RNA): template for protein synthesis

  • tRNA (transfer RNA): AA carrier

  • rRNA (ribosomal RNA): a component of ribosome for protein synthesis

  • hnRNA (heterogeneous nuclear RNA): precursor of mRNA

  • snRNA (small nuclei RNA): small RNAs for processing and transporting hnRNA

Unique features
Unique features

  • RNA is single stranded, in general.

  • RNA has self-complementary intrachain base paring.

  • The double helical regions of RNA are of the A-form.

  • RNA is susceptible to hydrolysis.

3 1 messenger rna
§ 3.1 Messenger RNA

  • mRNA is the template for protein synthesis, that is, to translate each genetic codon on mRNA into each AA in proteins. Each genetic codon is a set of three continuous nucleotides on mRNA.

  • mRNAs constitute 5% of total RNAs.

  • mRNAs vary significantly in life spans.

  • hnRNA is the precursor of mRNA.

mRNA maturation

  • hnRNA contains introns and exons.

  • Exons are the sequences encoding proteins, and introns are non-coding portions.

  • Splicing process of hnRNA removes introns and makes mRNA become matured.

  • The matured mRNA has special structure features, including 5’-cap and 3’-poly A tail.

5 cap

  • mRNA chain

5 cap addition1
5’-cap addition

  • Methylation can occur at different sites on G or A.

  • 5’-cap can be bound with CBP, benefiting transporting from nucleus to cytoplasm.

  • 5’-cap can be recognized bytranslation initiation factor.

  • It protects the 5’-end from exonucleases.

Poly a tail
Poly A tail

  • 20-200 adenine nucleotides at 3’ end

  • a un-translated sequence.

  • Related with mRNA degradation that begins with poly A tail shortening.

  • Associate with poly A tail binding proteins for protection



hnRNA splicing



3 2 transfer rna
§ 3.2 Transfer RNA

tRNA serves as an amino acid carrier to transport AA for protein synthesis.

  • tRNA is about 15% of total RNA.

  • tRNA is 65-100 nucleotides long.

  • There are at least 20 types of tRNA in one cell.

Structure of trna
Structure of tRNA

  • The overall structure is a cloveleaf, reversed L-shape structure.

  • There are three loops (DHU loop, anticodon loop, TψC loop),and four stems.

  • The 3-D structure is stabilized by hydrogen bonds of local intrachain base pairs on these stems.

Two key sites of tRNA

  • A tRNA molecule has an amino acidattachment site and a template-recognition site, bridging DNA and protein.

  • The template-recognition site is a sequence of three bases called the anticodon complementary to the mRNA codon.

Codon and anticodon

The anticodon on tRNA pairs with the codon on mRNA.

Amino acid attachment

  • The OH group at the 3' end of tRNA links covalently to an amino acid.

  • Only the attached AA becomes activated and capable of being transported.

Rare Bases

  • tRNA contains a high portion of unusual bases.

3 3 ribosomal rna
§ 3.3 Ribosomal RNA

rRNA provides a proper place for protein synthesis.

  • rRNA is the most abundant RNA in cells (>80%).

  • rRNA assembles with numerous ribosomal proteins to form ribosomes.


  • Ribosomes associate with mRNA to form a place for protein synthesis.

  • Ribosomes of eukaryotes and prokaryotes are similar in shapes and functions.

Components of ribosomes

Prokaryote Eukaryote

(E.coli) (Liver of mouse)

Smaller subunit 30s 40s

rRNA 16s 1542 nucleotides 18s 1874 nucleotides

proteins 21 40% of total weight 33 50% of total weight

Larger subunit 50s 60s

rRNA 23s 2940 nucleotides 28s 4718 nucleotides

5s 120 nucleotides 5.85s 160nucleotides

5s 120nucleotides

proteins 31 30% of total weight 49 35% of total weight

Ribosome of E. coli

Secondary structure of 18S rRNA

The secondary structure of rRNA has many loops and stems, which can bind ribosomal proteins to form an assembly for protein synthesis.

Section 4 physical and chemical properties of nucleic acids

Section 4Physical and Chemical Properties of Nucleic Acids

General properties
General properties

  • Acidity

    • Negative backbone

  • Viscosity

    • Concentration and aggregation effects

  • Optical absorption

    • UV absorption due to aromatic groups

  • Thermal stability

    • Disassociation of dsDNA (double-stranded DNA) into two ssDNAs (single-stranded DNA)

§ 4.1 UV Absorption

Application of OD260

  • Quantify DNAs or RNAs

    • OD260=1.0 equals to

      • 50μg/ml dsDNA

      • 40μg/ml ssDNA (or RNA)

      • 20μg/ml oligonucleotide

  • Determine the purity of nucleic acid samples

  • pure DNA: OD260/OD280 = 1.8

    • pure RNA: OD260/OD280 = 2.0

Transition of dsDNA to ssDNA

The absorbance at 260nm of a DNA solution increases when a dsDNA is melted into two single strands. The change is called hyperchromicity.

DNA melting

  • Melting curve: a graphic presentation of the absorbance of dsDNA at 260nm versus the temperature.

  • Melting temperature(Tm): the temperature at which the UV adsorption reaches the half of the maximum value, also means that about 50% of the dsDNA is disassociated into the single-stranded DNA.

Melting curve shift

Tm of dsDNA depends on its average G+C content. The higher the G+C content, the higher the Tm.

4 2 thermal stability
§ 4.2 Thermal stability

  • Dissociation of dsDNA into two ssDNAs is referred to as denaturation.

  • Denaturation can be partially and completely.

  • The nature of the denaturation is the breakage of H-bonds.

  • Denaturation is a common and important process in nature.

Denaturation of DNA

Extremes in pH or

high temperature

Cooperative unwinding

of DNA strands

Renaturation of dna
Renaturation of DNA

Two separated complementary DNA strands can rejoin together to form a double helical form spontaneously when the temperature or pH returns to the biological range. This process is called renaturation or annealing.

4 3 hybridization
§ 4.3 Hybridization

  • The ability of DNA to melt and anneal reversibly is extremely important.

  • An association between two different polynucleotide chains whose base sequences are complementary is referred to as hybridization.

  • The stability of the hybridized strand depends on the complementarydegree.

Two dsDNA molecules from different species are completely denutured by heating. When mixed and slowly cooled, complementary DNA strands of each species will associate and anneal to form normal duplexes.

  • Two ssDNAs, two ssRNAs, as well as one ssDNA and one ssRNA can also be hybridized.

  • Ionic strength, degree of complementary, temperature, as well as base composition, fragment length of nucleic acids will affect the hybridization.

  • It is a common phenomenon in biology, and has been used as a convenient techniques in medicine and biology.

Complementary hybridization

Target DNA detection

complementary hybridization

probe: …. TAGCTGAG …target: …. ATCGACTC …

  • mismatched hybridization

probe: …. TAGCTGAG …non-target: …. ATCAGCTC …


  • Gene structure and expression

  • Microarray or gene chip

  • mRNA separation

  • Gene diagnosis and therapy

  • PCR technique

Section 5 nuclease

Section 5Nuclease

Definition and classification

Nucleasesare enzymes that are able to hydrolyze phosphoester bonds and cleave DNA or RNA into fragments.

  • Deoxyribonuclease (DNase)- specially cleave DNARibonuclease (RNase) - specially cleave RNA


ExonucleasesThey can cleave terminal nucleotides either from 5’-end or from 3’-end, such as enzymes used in the DNA replication.Endonucleases They can cleave internally at either 3’ or 5’ side of a phosphate group, such as the restriction endonucleases used to construct the recombinant DNA.










  • Participate in DNA synthesis and repair, as well as RNA post-translational modification

  • Digest nucleic acids of food for better absorption

  • Degrade the invaded nucleic acids

  • Construct the recombinant DNA