Dr.

Umar Aliyu Abdullahi © 2023

BTG 202 InTroducTory BIoTechnoloGy

Cellular Organization at Molecular Level

Chemical Nature of Genetic Material

Nucleic acids are biopolymers consisting of smaller units (monomers) called nucleotides. Each
nucleotide consists of:
1. a sugar molecule (ribose or deoxyribose)
2. a mole of phosphate
3. a molecule of a nitrogenous base: Adenine (A), Guanine (G), Cytosine
(C), Thymine (T) and Uracile (U).
The nitrogenous base is covalently bonded to the number 1 carbon atom of one pentose
molecule. The phosphoric acid molecule is covalently bonded to the fifth carbon atom of the
pentose molecule.
There are two classes of nucleic acids: Deoxyribonucleic acid (DNA) and Ribonucleic acid
(RNA).
DNA; two forms of nitrogenous base are found in the DNA molecule: double-ring purines and
single-ring pyrimidines. The two purines of DNA are adenine and guanine, and the two
pyrimidines of DNA are the cytosine and thymine. A phosphate and a deoxyribose sugar
(pentose) molecule are common to all the four kinds of nitrogenous bases. The DNA molecule
consists of two helical polynucleotide chains wound around each other. In DNA A is
complementary to T and С is complementary to G. Complementary within each of these two
couples is based on the possibility of establishing hydrogen bonds that maintain the double-
helical structure. The two complementary polymers of nucleotides (called strands) that make
up a double helix are antiparallel (5' end faces 3' end and vice versa). That is, beginning at one
end of the molecule and progressing toward the other, successive nucleotides of the same chain
are joined together by 3'→5' phosphodiester bonds, whereas the complementary nucleotides
of the other chain are joined by 5'→3' phosphodiester bonds. The exact arrangement of purine-
pyrimidine matches is specific for every species of organism.
RNA; They differ from DNA by the fact that the pentose molecule is ribose instead of
deoxyribose in DNA and the nitrogenous base uracil instead of thymine. RNA synthesis is
directed by specific segments of DNA (genes). Generally, only one strand (not chosen
randomly) is read in transcription. RNA is divided into several types: Ribosomal RNA (r-RNA,
 Dr. Umar Aliyu Abdullahi © 2023

representing up to 85 % of total RNA); Messenger RNA (m-RNA, representing up to 5 % of

total RNA); Transfer RNA (t-RNA, representing about to 10 % of total RNA).
Gene
A gene is the basic physical and functional unit of heredity. Genes are components of genetic
material and are thus units of inheritance. Genes are made up of DNA. Some genes act as
instructions to make molecules called proteins while other do not. Gene is a segment of DNA
that codes for one polypeptide, one t-RNA or one ribosomal RNA (r-RNA) molecule.

Central Dogma of Molecular Biology

The flow of genetic information from DNA to RNA by the process of transcription and from
RNA to proteins by the process of translation called the central dogma of molecular biology.

Transcription
The process of copying the genetic information of the DNA to mRNA (messenger RNA) by RNA
polymerase is called transcription. The DNA strand which is transcribed is called template
strand. The mRNA thus produced is complementary to the template stand and identical to the
sense strand.
Stages of Transcription
Initiation
Transcription is catalysed by the enzyme RNA polymerase, which attaches to and moves along
the DNA molecule until it recognises a promoter sequence. This area of DNA indicates the
starting point of transcription, and there may be multiple promoter sequences within a DNA
molecule. Transcription factors are proteins that control the rate of transcription; they too bind
to the promoter sequences with RNA polymerase. Once bound to the promoter sequence, RNA
polymerase unwinds a portion of the DNA double helix, exposing the bases on each of the two
DNA strands.
Elongation
One DNA strand (the template strand) is read in a 3′ to 5′ (three-prime to five-prime) direction,
and so provides the template for the new mRNA molecule. The other DNA strand is referred
to as the coding strand. This is because its base sequence is identical to the synthesised mRNA,
except for the replacement of thiamine bases with uracil. RNA polymerase uses incoming
ribonucleotides to form the new mRNA strand. It does this by catalysing the formation of
phosphodiester bonds between adjacent ribonucleotides, using complementary base pairing (A
 Dr. Umar Aliyu Abdullahi © 2023

to U, T to A, C to G, and G to C). Bases can only be added to the 3′ end, so the strand elongates
in a 5’ to 3’ direction.
Termination
Elongation continues until the RNA polymerase encounters a stop sequence. At this point,
transcription stops, and the RNA polymerase releases the DNA template.

Pre-translational mRNA processing

The newly synthesized mRNA remaining inside the nucleus is called hn-RNA (heterogeneous
nuclear RNA). This hn-RNA is processed addition of methylated guanine cap at the 5’ end
and by addition of several adenines (poly –A tail) at 3’ end. This capping and addition of poly
A-tail protects the mRNA from degradation by enzymes called RNase. This molecule
undergoes severe alterations to remove non-coding parts called introns, leaving only the coding
parts or exons to produce the mRNA. This mRNA is about 25% of the original length. The
mRNA is then transported from the nucleus to the ribosomes in the cytoplasm for protein
synthesis.
Splicing allows the genetic sequence of a single pre-mRNA to code for many different proteins,
conserving genetic material. This process is sequence-dependent and occurs within the
transcript. It involves:

 Removal of introns (non-coding sequences) via spliceosome excision

 Joining together of exons (coding sequence) by ligation
Note:
INTRON refers to the non-coding region of the eukaryotic genes that are transcribed into
mRNA. They are removed by splicing of RNA.
EXON refers to the region of DNA that codes for a protein. In eukaryotes, the exons are
separated by many introns.
Translation
The process by which the nucleotides sequence present in the mRNA is translated into amino
acid sequence is called as translation. During translation, an mRNA sequence is read using the
genetic code, which is a set of rules that defines how an mRNA sequence is to be translated
into the 20-letter code of amino acids, which are the building blocks of proteins. The genetic
code is a set of three-letter combinations of nucleotides called codons, each of which
 Dr. Umar Aliyu Abdullahi © 2023

corresponds with a specific amino acid or stop signal. Translation of an mRNA molecule by
the ribosome occurs in three stages: initiation, elongation, and termination.
During initiation, the small ribosomal subunit binds to the start of the mRNA sequence. Then
a transfer RNA (tRNA) molecule carrying the amino acid methionine binds to what is called
the start codon of the mRNA sequence. The start codon in all mRNA molecules has the
sequence AUG and codes for methionine. Next, the large ribosomal subunit binds to form the
complete initiation complex.
Elongation stage, the ribosome continues to translate each codon in turn. Each corresponding
amino acid is added to the growing chain and linked via a bond called a peptide bond.
Elongation continues until all of the codons are read.
Termination stage occurs when the ribosome reaches a stop codon (UAA, UAG, and UGA).
Since there are no tRNA molecules that can recognize these codons, the ribosome recognizes
that translation is complete. The new protein is then released, and the translation complex
comes apart.
Codon: A specific DNA or RNA sequence of three nucleotides corresponding to a particular
amino acid.

Gene expression
Gene expression is the process by which the information encoded in a gene is turned into a
function. This mostly occurs via the transcription of RNA molecules that code for proteins or
non-coding RNA molecules that serve other functions. It consists of two major steps:
transcription and translation. Together, transcription and translation are known as gene
expression. During the process of transcription, the information stored in a gene's DNA is
passed to a similar molecule called RNA (ribonucleic acid) in the cell nucleus.

Regulation of Gene Expression in Prokaryotes

Regulation of gene expression refers to the turning on and turning off of the genes at
appropriate time and to the desired level. It is important for proper growth and development.
The gene expression may be regulated during transcription, mRNA processing, translation, and
involve posttranslational modifications.
An Operon is a group of structural genes whose transcription is regulated by the action of a
regulator gene ‘r’, a promoter gene ‘p’ and an operator gene ‘o’.
 Dr. Umar Aliyu Abdullahi © 2023

Regulation of Operon

There are two basic categories of gene regulation: negative and positive.
1. In negative regulation, an inhibitor which is bound to the DNA must be removed for
transcription to occur. In this case the transcription is normally on and must be turned off. It is
called repressible operon.
2. In positive regulation, an activator has to bind to the DNA for transcription to occur. In this
case, the transcription is normally off and must be turned on. It is called as inducible operon.

Regulation of Gene Expression in Eukaryotes

Gene expression in eukaryotic cells is regulated by repressors as well as by transcriptional
activators (Some transcription factors activate transcription, helping the general transcription
factors and/or RNA polymerase bind to the promoter). Like their prokaryotic counterparts,
eukaryotic repressors bind to specific DNA sequences and inhibit transcription. In some cases,
eukaryotic repressors simply interfere with the binding of other transcription factors to DNA.
For example, the binding of a repressor near the transcription start site can block the interaction
of RNA polymerase or general transcription factors with the promoter, which is similar to the
action of repressors in bacteria. Other repressors compete with activators for binding to specific
regulatory sequences. Some such repressors contain the same DNA-binding domain as the
activator but lack its activation domain. As a result, their binding to a promoter or enhancer
blocks the binding of the activator, thereby inhibiting transcription.