REPLICATION

 After Watson and Crick deduced the three dimensional structure of DNA, they
immediately inferred its mechanism of replication. The double
 helical model of DNA immediately suggested a mechanism for the replication
of DNA.
 Watson and Crick proposed that one of the strands of each daughter DNA
molecule is newly synthesized, whereas the other is derived from the parent
DNA molecule. This distribution of parental atoms is called semi-
conservative.
 The 2 strands of DNA helix come apart when the H bonds between its paired
bases are disrupted. This can be done by heating a solution of DNA or by
adding acid or alkali to ionize its nitrogenous bases.
 REPLICATION
 Replication is a process by which DNA molecules produce identical copies of
themselves.
 Characteristics of DNA replication:
 * Semiconservative – each daughter DNA contains one strand from the
 parent DNA.
 * Bidirectional – occurs in two directions about the origin of replication.
 - occurs in the 5’ - 3’ direction
* Discontinuous synthesis – the lagging strand is first made as short chains
3. (Okazaki fragments) which are subsequently linked to form long chains.
* Newly synthesized strand is complementary to the parent strand following
• the specific base pairing, thus:
• 3’ CGTACCTTA 5’ parent strand
• 5’ GCATGGAAT 3’ newly synthesized strand
 REPLICATION
The unwinding of DNA molecule during replication is facilitated by
different enzymes:
 A. Primase synthesizes a short strand of RNA (on the DNA template) which
 act as a primer in initiating DNA polymerization.

 B. DNA Polymerase sequentially polymerizes nucleotides to form a new DNA

 strand. DNA polymerases also have exonuclease properties – the ability to
 hydrolyze and remove improperly paired terminal nucleotides.

 C. DNA Ligase could catalyze the formation of a phosphodiester bond

 between two DNA chains. The DNA ligase catalyzes the joining of two
 DNA chains that are part of a double helical molecule. This enzyme
 requires a free OH group at the 3’ end of one DNA chain and a phosphate

 REPLICATION

 group a the 5’ end of the other DNA strand. The DNA chains
 joined by the DNA ligase must be part of a double helical
DNA molecule.
 This joining process is essential for the normal synthesis of
DNA, for repair of damaged DNA and for the splicing of DNA
 chains in genetic recombination. It also connects newly
 formed fragments (Okazaki fragments) of DNA with one
 another to form a single chain.
 D. Endonuclease

 Endonucleases can hydrolyze connections between those nucleotides that lie

 between those nucleotides that lie within the central area of the nucleotide
 chain (as opposed to exonucleases , which hydrolyze terminal positions. A
 specific endonuclease can help remove a segment of DNA rendered
defective
 by UV light. The latter can damage DNA by making two adjacent thymine
 bases fuse covalently. The endonuclease makes a nick in the DNA strand on
 one side of the defective thymine dimer. An exonuclease removes the
 damaged DNA. A DNA plolymerase adds replacement nucleotides. DNA
 ligase reattaches the newly synthesized DNA to the original strand.
 REPLICATION
 Interplay of Molecules in Replication
 At replication fork, both strands of parental DNA serves as templates for the
synthesis of new DNA. Recall that parental strands are antiparallel. Hence,
the overall direction of DNA synthesis must be 5’ ---- 3’ for one daughter
strand and 3’ ----- 5’ for the other. However, all known DNA polymerases
synthesize DNA in the 5’ ------- 3’ direction, but not in the 3’ ------- 5’
direction.
 A large proportion of newly synthesized DNA exists as small fragments. These
are called Okazaki fragments. As replication proceeds, these fragments
become covalently joined by DNA ligase to form one of the daughter strands.
The strand formed from Okazaki fragments is termed the lagging strand,
whereas the one synthesized with few or no interruptions is the leading
strand. Both Okazaki fragments and leading strand are synthesized in the
 5’ ----- 3” direction.
 REPLICATION
 Requirements of DNA Replication:
 Template - provides sequence information
 Primer - provides free ‘OH to which deoxynucleotides are added
 Precursors – Deoxynucleotide triphosphate (DNTPs); examples are dATP
 (deoxy adenosinetriphosphate), dGTP (deoxy guanosinetriphosphate), etc.

 Stages of DNA Replication:

 1. Initiation
 2. Elongation and
 3. Termination
 THE RNAS
 Molecules of RNA make up 5 – 10 % of the total weight of the cell. The
nucleotides of RNA contain ribose instead of deoxyribose, and the base uracil
instead of thymine. Unlike DNA, RNA is not double stranded. It consists of a
single strand of nucleic acid. There are three distinct types of RNAs found in the
cell: messenger RNA (mRNA), ribosomal RNA (rRNA) and transfer RNA (tRNA)
 Messenger RNA (mRNA)
 -mRNA is synthesized in the nucleus of the cell, and contains the four
bases A, C, G and U, It is synthesized on one strand of a DNA helix, so it
will have a sequence of bases complementary to that of the DNA. mRNA
is not a stable molecule and is synthesized by the cell whenever it is
needed. After being synthesized, mRNA molecule will migrate to the
cytoplasm of the animal cell. There it will serve as a pattern for the
sequencing of amino acids in the synthesis of proteins in the ribosomes.
 -It carries genetic messages from DNA to ribosomes where protein
 synthesis occurs.
 THE RNAS

 Ribosomal RNA (rRNA)

 rRNA is also synthesized in the cell from a DNA template. It migrates
to the cytoplasm of the cell where with protein it forms the
ribosomes.
 Transfer RNA (tRNA)
 tRNA is the smallest of the RNA molecules. It is synthesized in the nucleus of
the cell and each is specifically designed for a particular amino acid. A tRNA
molecule becomes charged when a specific amino acid is joined to the
terminal adenine nucleotide present on each RNA polynucleotide chain. The
tRNA then carries this amino acid to the ribosomes, where the amino acids
are used in protein synthesis.
 TRANSCRIPTION
 Characteristics of transcription:
 * Only one strand is transcribed. template strand or antisense strand – which
 is complementary to the strand containing the gene – coding strand or sense
 strand
 * transcription happens via specific base pairing – C with G, A with U
 * mRNA transcribed is a copy of the coding strand; differ only in having U
 instead of T
 Example: 5’ C T A A T G G T 3’ DNA coding strand
 3’ G A T T A C C A 5’ DNA template strand
 5’ C U A A U G G U 3’ Transcribed mRNA

 TRANSCRIPTION
 Stages of Transcription:
 1. Initiation
* RNA POLYMERASE BINDS
• DNA STRANDS SEPARATE
• First nucleotides are bound
• Second nucleotides are bound
• First phosphodiester bond is formed
2. Elongation
* Addition of further nucleotides
* Formation of phosphodiester bond
3, Termination
• Synthesis stops
• RNA is released
• DNA helix reforms
 TRANSLATION
 Translation or Protein Synthesis
* specific ribonucleotide sequences in mRNA form a message that determines
• the order in which different amino acid residues are to be joined
* Message carried by mRNA through codons is translated into a specific amino
• acid sequence characterizing a specific protein

• Codon
• * a sequence of three ribonucleotides specific for a given amino acid
• * since 43 = 64, there are 64 triplets for the four bases in RNA; 61 code for
• specific amino acids and 3 code for chain termination
• (stop/termination codons)
• * the AUG codon aside from coding for the amino acid methionine, can
•

 *
 TRANSLATION
• function as the start/initiator codon when it occurs as the first codon in the
• mRNA
• Example: UACCGACUCAUUUAC
• Tyr Arg Leu Ile Tyr

• Genetic Code
• * gives the assignment of the 64 mRNA codons to specific amino acids or
• stop signals

• *
 TRANSLATION
 Five major stages

 1. Activation stage
 2. Initiation
 3. Elongation
 4. Termination and release
 5. Folding and processing
REFERENCES:
 Biochemistry by Campbell
 Lehninger’s Principles of Biochemistry
 UPOU manual in Biochemistry
 Internet sources
 MUTATION
 Mutation is any chemical or physical change in the DNA molecule which results
in the synthesis of a protein having an altered amino acid sequence.
 - Some forms of mutation are beneficial to an organism, but more will be
detrimental to varying degrees.

Ways by which a gene can be changed or mutated.

1. substitution of one base pair for another of several pairs
2. the deletion of one or more base pairs
3. the insertion of one or more base pairs