MODULE –VI (Information Transfer)

Genetic material serves three crucial functions:

1. Genotypic function: It transmits genetic information from one generation to another.
2. Phenotypic function: It directs growth and differentiation. It also leads to the development
of offspring into reproductive traits.
3. Evolutionary function: Mutations enable organisms to respond to environmental changes.

DNA as a Genetic Material

The notion that protein was the genetic material initially remained strong but was
eventually overturned by the results of two important scientific experiments: A) Griffith’s
Experiment and B) Hershey Chase experiment. DNA stores genetic information; it encodes
the sequence of amino acid residues in proteins using the genetic code, triplet code.

A)Transforming principle: Griffith’s Experiment

In 1928, Frederick Griffith conducted an experiment specifying the capability of bacteria

to transfer genetic information. He studied the difference between pathogenic and non-
pathogenic strains of pneumonia-causing bacteria, Streptococcus pneumonia. The pathogenic
strain (Smooth- S strain) was surrounded by a capsule and caused pneumonia, whereas the non-
pathogenic strain (Rough-R strain) did not have a capsule and did not cause pneumonia.
Griffith injected these two strains of bacteria into mice. The S strain killed the mice, while
the R strain did not. He further found that when heat-killed S strain was injected into mice, it
did not cause pneumonia. Later, he combined the heat-killed S strain with the live R strain and
injected the mixture into a mouse; the mouse developed pneumonia and died. He determined
that the non-pathogenic R strain could be transformed into the pathogenic S strain if the remains
of the dead virulent strain are available to the living non-virulent strain and termed it a
transforming principle.
B) Hershey-Chase Experiment
They worked with viruses that infect bacteria called bacteriophages. The bacteriophage
attaches to the bacteria and its genetic material then enters the bacterial cell. The bacterial cell
treats the viral genetic material as if it was its own and subsequently manufactures more virus
particles.
They grew some viruses on a medium that contained radioactive phosphorus and some
others on medium that contained radioactive sulfur. Viruses grown in the presence of
radioactive phosphorus contained radioactive DNA but not radioactive protein because DNA
contains phosphorus but protein does not. Similarly, viruses grown on radioactive sulfur
contained radioactive protein but not radioactive DNA because DNA does not contain sulfur.

Radioactive phages were allowed to attach to E. coli bacteria. Then, as the infection
proceeded, the viral coats were removed from the bacteria by agitating them in a blender. The
virus particles were separated from the bacteria by spinning them in a centrifuge.
Bacteria which was infected with viruses that had radioactive DNA were radioactive,
indicating that DNA was the material that passed from the virus to the bacteria. Bacteria that
were infected with viruses that had radioactive proteins were not radioactive. This indicates
that proteins did not enter the bacteria from the viruses. DNA is therefore the genetic material
that is passed from virus to bacteria.

Organization of Genetic Material

DNA is protected and packaged in very specific ways. In addition, DNA molecules can
be very long. Stretched end-to-end, the DNA molecules in a single human cell would come to
a length of about 2 meters. Thus, the DNA for a cell must be packaged in a very ordered way
to fit and function within a structure (the cell).
The vast majority of an organism’s genome( entre set of DNA instructions found in a cell) is
organized into the cell’s chromosomes, which are discrete DNA structures within cells that
control cellular activity. Eukaryotic chromosomes are housed in the membrane-bound nucleus,
The nucleus of a eukaryotic cell, it is surrounded by a nuclear membrane.

Most prokaryotes contain a single, circular chromosome that is found in an area in the
cytoplasm called the nucleoid.
The nucleoid : It is an irregularly-shaped region within the cell of a prokaryote that contains
all or most of the genetic material. The length of a genome widely varies, but generally is at
least a few million base pairs.

(Each chromosome is made of protein and a single molecule of deoxyribonucleic acid (DNA)).

Organization of Prokaryotic Chromosomes

The size of the genome in one of the most well-studied prokaryotes, Escherichia coli, is 4.6
million base pairs, which would extend a distance of about 1.6 mm if stretched out. The DNA
is twisted beyond the double helix in what is known as supercoiling. Some proteins are known
to be involved in the supercoiling; other proteins and enzymes help in maintaining the
supercoiled structure.
Chromosomes in bacteria and archaea are usually circular and typically contains only a single
chromosome within the nucleoid. Because the chromosome contains only one copy of each
gene, prokaryotes are haploid.
The nucleoid is composed of DNA in association with a number of DNA-binding proteins
(histone-like proteins) that help it maintain its structure. The isolated nucleoid contains 80%
DNA, 10% protein, and 10% RNA by weight.
There are four major families of bacterial histone-like proteins(but not histones):
1- Histone-like nucleoid structuring protein (H-NS)
2- Heat unstable protein (HU)
3- Factor for inversion stimulation (FIS)
4- Integration host factor (IHF)
Proteins or nucleoid proteins or nucleoid-associated proteins (NAPs) and are distinct from
histones of eukaryotic nuclei.
In contrast to histones, the DNA-binding proteins of the nucleoid do not form
nucleosomes, in which DNA is wrapped around a protein core. Instead, these proteins often
use other mechanisms to promote compaction such as DNA looping. The most studied NAPs
are HU, H-NS, FIS, and IHF that organize the genome by driving events such as DNA bending,
bridging, and aggregation.

Fig- Structure of bacterial chromosome

Organization of Eukaryotic Chromosome

Eukaryotic chromosomes are typically linear, and eukaryotic cells contain multiple distinct
chromosomes. Many eukaryotic cells contain two copies of each chromosome and, therefore,
are diploid.
During DNA packaging, DNA-binding proteins called histones perform various levels of DNA
wrapping and attachment to scaffolding proteins. The combination of DNA with these attached
proteins is referred to as chromatin. (Chromatin is a complex of DNA and proteins that
forms chromosomes within the nucleus of eukaryotic cells. Nucleosome consist of DNA
wrapped around histone proteins and is the basic repeating unit of Chromatin)
In eukaryotes, the packaging of DNA by
histones may be influenced by
environmental factors that affect the
presence of methyl groups on certain
cytosine nucleotides of DNA. The influence
of environmental factors on DNA
packaging is called epigenetics.
Each species of plants and animals has a set
number of chromosomes. Eg. Humans
have 46 chromosomes while a rice plant has
12.
The functions of chromatin are:
- To package DNA into a smaller volume to
fit in the cell.
- To strengthen the DNA to allow mitosis
and meiosis.
Fig- Hierarchical levels of DNA organization
Histones

Histones are highly basic proteins found in eukaryotic cell nuclei that pack and order the DNA
into structural units called nucleosomes. Five major families of histones exist: H1, H2A, H2B,
H3, and H4. Histones H2A, H2B, H3 and H4 are known as the core histones, while histones
H1/H5 are known as the linker histones. All four of the core histones contain between 20 and
25% of lysine and arginine.

Out side the nucleosome structure there are other type of proteins called non histone protein.
A variety of non-histone proteins also bind to DNA to affect chromatin structure and exert
epigenetic control on gene expression.

Struture -
The basic structural unit of DNA packaging in eukaryotes are called nucleosomes. Each
nucleosome is made of DNA wrapped around eight histone proteins that function like a spool
and are called a histone octamer .
Histones are a family of basic proteins that associate with
DNA in the nucleus and help condense it into chromatin.
Nuclear DNA does not appear in free linear strands; it is
highly condensed and wrapped around histones in order to
fit inside of the nucleus and take part in the formation of
chromosomes.
Each histone octamer is composed of two copies each of the
histone proteins H2A, H2B, H3, and H4. The chain of
nucleosomes is then wrapped into a 30 nm spiral called a
solenoid, where additional H1 histone proteins are
associated with each nucleosome to maintain the
chromosome structure. Each nucleosome attached with
followed one by linker DNA ( 20-60 BP) thus total DNA
warp around it which will be protected from digestion with
micrococcal endonuclease. The fifth histone H1 usually
exist out side the core (in the binding region between
nucleosome and another).
Levels of DNA Packing-
-First level twisting or super coiling of
DNA molecules
- Second level warping of DNA around
histons.
- Formation of folds or zig-zag by H1
histone and the linker (other benefits of
linker create elasticity and flexibility to
chromatin beside binding two adjacent
nucleosome)
- Formation of (30 nm) fibers and solenoid
model by collecting each 6 nucleosome
together.

Gene structure - The genetic material DNA is organized into basic physical and functional
units of heredity called genes. A gene is a region of DNA encodes the synthesis of the gene
product, RNA, and protein. Basically, genes in the DNA encode proteins that carry all the
functions inside the cell, including metabolizing nutrients and synthesis of new cellular
constituents.

MOLECULAR BASIS OF INFORMATION TRANSFER:

The genetic information stored in the nucleotide sequence of DNA serves two purposes:
1. It provides the information inherited by daughter cells or offspring.
2. It is the source of information for the synthesis of all protein molecules of the cell and
organism. It is called the central dogma of biological information. The Central Dogma explains
the flow of genetic information and was proposed in 1958 by Francis Crick. It refers to the
intricate base-to-base transfer of genetic information from nucleic acids to proteins via RNA.
The expression of genes, meaning the synthesis of proteins, constitutes two major steps:
● Transcription - It is the synthesis of
single-stranded RNA molecules from
double-stranded DNA, basically a
transfer of genetic information from
DNA to RNA.
● Translation - It is the process of
reading the mRNA sequence as the
genetic code that transfers the
information stored in the DNA into the
amino acid sequence of proteins.
Concept of Genetic Code :
The genetic code is the set of rules by which information encoded in DNA is translated into
proteins. The sequence of nucleotide bases (A, T, C, G) in DNA determines the sequence of
amino acids in proteins.

General features of genetic code:

1. The genetic code is a triplet code called a codon, meaning a triplet of nucleotides
codes for a specific amino acid or give signal to stop during protein synthesis.
Example: The codon "AUG" codes for the amino acid methionine (also the start
codon).

2. Each triplet specifies a specific amino acid; thus, the coding is unambiguous(show
specificity). A given codon stands for only 1 Amino acid. Eg- UGG always codes
for Tryptophan.

3. The code consists of start and stop signals, which are required to initiate and
terminate translation. Three codons out of 64 do not code for any amino acids and are
known as stop codons or non-sense codons (UAA, UAG and UGA), while the code
AUG acts as a start codon and codes for methionine.

4. Once the translation of mRNA commences, codons are read consecutively with no
gaps in between. Thus, the code is said to be comma less.

5. The code is degenerate. A particular amino acid can be specified by more than one
triplet codon.
 6. The code is non-overlapping. (Codes are read one after another)

7. The genetic code is nearly universal, meaning the code remains the same for almost
all organisms with few exceptions.

Degeneracy/Redundancy of genetic code:

The genetic code is said to be degenerate because multiple codons can encode the same amino
acid. This redundancy arises because there are 64 possible codons (43 combinations of the bases
A, T, C, G), but only 20 standard amino acids are used in proteins. This means a particular
amino acid can be specified by more than one triplet codon. The difference between them lies
at the third position, while the first two bases of the codon determine specificity. This is true
for 18 of the 20 amino acids. The different codons for a particular amino acid are synonymous.
The Wobble hypothesis explains the degeneracy of the codons.

Examples of Degeneracy:

- Leucine is encoded by six different codons: "CUU", "CUC", "CUA", "CUG", "UUA",
"UUG".

- Arginine is encoded by six different codons and Glycine has 4 codons.

Universality of Genetic Code:

The genetic code is nearly universal across all living organisms, from bacteria to humans
(Except few - These exceptions have been found in the genome of protozoans, mycoplasma,
and mitochondria. For example, from bacteria to humans, UGA is a stop codon, but in
mitochondrial DNA and Mycoplasma capricolum, it codes for tryptophan. Another variation
occurs in ciliated protist, where both the termination codons, UAG and UAA, codes for
glutamine)
This means that:

 The same codons specify the same amino acids in nearly all organisms.
  The universality of the genetic code provides evidence for the common origin of life on
Earth and is fundamental in biotechnology, as genes from one organism can be
expressed in another organism.

Exceptions to genetic code-