CHAPTER 6

MOLECULAR BASIS OF
INHERITANCE
RNA
RNA though it also acts as a genetic
materials1 in some viruses, mostly functions
as a messenger2 (mRNA).
RNA has
additional roles as well. It functions as
adapter3 (tRNA), structural4 molecule
(rRNA), and in some cases as a catalytic5
molecule (riboswitches, Ribozymes) or a
regulatory6 molecule (snRNA like miRNA
and siRNA).
DNA
DNA is a long polymer of
deoxyribonucleotides.
The length of DNA is usually defined as
number of nucleotides or base pairs.

Bacteriophage 174 has 5386

nucleotides,
Bacteriophage lambda has 48502
base pairs
Escherichia coli has 4.6 106 bp,
Haploid content of human DNA is 3.3
109 bp.

STRUCTURE OF DNA
A nucleotide has three components a
nitrogenous base, a pentose sugar (ribose in
case of RNA, and deoxyribose for DNA), and
a phosphate group.
There are two types of nitrogenous bases
Purines (Adenine and Guanine), and
Pyrimidines (Cytosine, Uracil and Thymine).
A nitrogenous base is linked to the pentose
sugar through a N-glycosidic linkage to
form a nucleoside, such as adenosine,
guanosine, cytidine and uridine or its DNA
counterparts deoxyadenosine,
deoxyguanosine, deoxycytidine and
deoxythymidine.

A phosphate group is linked to 5'-OH of a

nucleoside through phosphoester
linkage, to form a nucleotide
Two nucleotides are linked through 3'-5'
phosphodiester linkage to form a
dinucleotide.
Polynucleotides are formed through these
phosphodiester linkages.
In RNA, every nucleotide residue has an
additional OH group present at 2'position in the ribose.
Also, in DNA the methylated form of
uracil, known as thymine (5-methyl
uracil) is found at the place of uracil.
DNA as an acidic substance Nuclein present
in nucleus was first identified by Friedrich
Meischer in 1869.
In 1953 that James Watson and Francis Crick,
based on the X-ray diffraction data produced
by Maurice Wilkins and Rosalind Franklin,
proposed the famous Double Helix model for
the structure of DNA.
Main Hallmark of the proposition
The unique property called complementary
base-pairing (discovered based on the
observation of Erwin Chargaff that for a
double stranded DNA, the ratios between
Adenine and Thymine and Guanine and
Cytosine are constant and equals one.)
This property allowed each of the two
strands of parental DNA to act as a template
for synthesis of new daughter strands that
are identical to the parental DNA molecule.
Because of this, the genetic implications of
the structure of DNA became very clear.
Key features of the double-helix structure

It is made of two polynucleotide chains,

where the backbone is constituted by
sugar-phosphate, and the bases project
inside.
The two chains have anti-parallel polarity.

The bases in two strands are paired

through hydrogen bonding. Adenine forms
two hydrogen bonds with Thymine, while
Guanine forms three H-bonds with
Cytosine. This as well as the fact that only
a uniform distance between the two
strands of helix is energetically feasible
makes sure that only a purine comes
opposite to a pyrimidine. This is also the
molecular reason for complementarity.
The two chains are coiled in a righthanded fashion.
The pitch of the helix is 3.4 nm and there
are roughly 10 bp in each turn. So, the
distance between each base-pair is
approximately equal to 0.34 nm. The
diameter of the B-DNA helix is roughly
2nm.

Figure: The central dogma of molecular

biology (extended)

PACKAGING OF THE DNA HELIX

In prokaryotes, such as, E. coli, though they
do not have a defined nucleus, the DNA is
not scattered throughout the cell. DNA
(being negatively charged) is held with some
proteins (that have positive charges) in a
region termed as nucleoid. The DNA in
nucleoid is organised in large loops held by
proteins.
In eukaryotes, this organisation is much
more complex. There is a set of positively
charged, basic proteins called histones. A
protein acquires charge depending upon the

abundance of amino acid residues with

charged side chains. Histones are rich in the
basic amino acid residues lysine and
arginine. Both the amino acid residues carry
positive charges in their side chains.
Histones are organised to form a unit of eight
molecules called as histone octamer. The
negatively charged DNA is wrapped around
the positively charged histone octamer to
form a structure called nucleosome. A typical
nucleosome contains 200 bp of DNA helix.
Nucleosomes constitute the repeating unit of
a structure in nucleus called chromatin,
thread-like stained (coloured) bodies seen in
nucleus. The Nucleosomes in chromatin are
seen as beads-on-string structure when
viewed under electron microscope.
The beads-on-string structure in chromatin is
packaged to form chromatin fibres that are
further coiled and condensed at metaphase
stage of cell division to form chromosomes.
The packaging of chromatin at higher level
requires additional set of proteins that
collectively are referred to as Non-histone
Chromosomal (NHC) proteins.
In a typical nucleus, some region of
chromatin are loosely packed (and stains
light) and are referred to as euchromatin.
The chromatin that is more densely packed
and stains dark are called as
Heterochromatin. Euchromatin is said to be
transcriptionally active chromatin, whereas
heterochromatin is inactive.
THE SEARCH FOR GENETIC MATERIAL
By 1926, the quest to determine the
mechanism for genetic inheritance had
reached the molecular level. Previous
discoveries by Gregor Mendel, Walter Sutton,
Thomas Hunt Morgan and numerous other
scientists had narrowed the search to the
chromosomes located in the nucleus of most
cells. But the question of what molecule was
actually the genetic material had not been
answered.
Transforming Principle

In 1928, Frederick Griffith, in a series of

experiments with Streptococcus
pneumoniae, witnessed a miraculous
transformation in the bacteria (a literal
change in the physical form of a living
organism).

They purified biochemicals (proteins, DNA

and RNA) from the heat-killed S cells to see
which ones could transform live R cells into S
cells. They discovered that DNA alone from S
bacteria caused R bacteria to become
transformed.

There are two kinds of strains of

Streptococcus pneumoniae bacteria: smooth
shiny strain (S) with mucous polysaccharide
coat (which make them virulent) and rough
strain (R) which lacks the coat.

For greater credibility, they used proteindigesting enzymes (proteases), RNAdigesting enzymes (RNases) and DNAdigesting (DNases) and found that only
DNases inhibited transformation, suggesting
that the DNA is the hereditary material once
again.

Observations:

Mice infected with the S strain

(virulent) die from pneumonia
infection

Mice infected with the R strain did not

die.

Heat-killed S strain bacteria injected

into mice did not kill them either.

But, when he injected a mixture of

heat-killed S and live R bacteria, the
mice died. Moreover, he recovered
living S bacteria from the dead mice.

The unequivocal proof: the Genetic

Material is DNA
This proof came in 1952 from the blender
experiments of Alfred Hershey and Martha
Chase. 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. The bacteriophage had two
components: a protein coat and a DNA.

Conclusion: the R strain bacteria had

somehow been transformed by the heatkilled S strain bacteria. Some transforming
principle, transferred from the heat-killed S
strain, had enabled the R strain to synthesize
a smooth polysaccharide coat and become
virulent. This must be due to the transfer of
the genetic material.
However, the biochemical nature of genetic
material was not defined from his
experiments.
Biochemical Characterization of
Transforming Principle
Oswald Avery, Colin MacLeod and Maclyn
McCarty were first experimenters behind the
determination of the biochemical nature of
Griffith's transforming principle.

Hershey and Chase worked to discover

whether it was protein or DNA from the
viruses that entered the bacteria.
They grew some viruses on a medium that
contained radioactive phosphorus and some
others on medium that contained radioactive
sulfur.

It was known that DNA contains phosphorous

and no sulfur while protein contained sulfur
but no phosphorous.
So, viruses/bacteriophages grown in the
presence of radioactive phosphorus
contained radioactive DNA but not
radioactive protein and similarly, viruses
grown on radioactive sulfur contained
radioactive protein but not radioactive DNA.
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. The bacteriophage coat was
frothed up in the supernatant while the
infected bacteria settled as sediments.
Radioactivity was detected in the
supernatant for the culture grown on
radioactive sulphur while the batch grown on
radioactive phosphorous showed
radioactivity as associated with the
sediment.
This result clearly indicated that the genetic
material passed on from virus to bacteria is
the DNA.

Though protein was displaced by DNA as the

true candidate for the genetic material, it
subsequently became clear that rarely, in
some viruses, RNA is the genetic material
not the DNA e.g. Tobacco Mosaic viruses and
QB bacteriophage.
Why does the DNA act as the predominant
genetic material, whereas the RNA mostly
performs the dynamic functions of a
messenger and an adapter? or to paraphrase
the question, why does the DNA seem a
better molecule to build a genome when
compared to RNA?
A molecule that can act as a genetic material
must fulfill the following criteria:

It should be able to generate its replica.

It should be chemically and structurally

stable.

It should provide the scope for slow

changes (mutation) that are required for
evolution.

It should be able to express itself in the

form of 'Mendelian Characters.

Since, the feature of complementary base

pairing applies to both DNA and RNA, both
are capable of replication.
But when the stability criterion is looked at, it
becomes clear that DNA supersedes RNA in
terms of both physical and chemical stability.
The naturally occurring double stranded
nature of DNA confers it great stability in
terms of physical influences like heat or
mechanical stress. Chemically, two key
reasons associated with the biochemical
nature of the RNA makes it more labile and
reactive.
1. The 2-hydroxyl group makes the RNA
susceptible to base-catalyzed hydrolysis and
hence easier degradation.

Properties of Genetic Material (DNA

versus RNA)

2. The replacement of Uracil by its

methylated form, Thymine grants selfrepair ability to DNA molecules. [Cytosine

deaminates at a perceptible rate to become

Uracil in all nucleic acids. But this change
can have deleterious effects as far as the
genomic information is concerned. Since
Cytosine base pairs with Guanine while
Uracil base pairs with Adenine, every such
change becomes equivalent to a pointmutation. But in DNA, a repair mechanism
involving the enzyme, Uracil DNA glycolase,
hydrolyses Uracil residues (and replaces it
with Cytosine) while keeping its methylated
form, Thymine, unscathed. In fact, the
methyl group on thymine is a tag that
distinguishes thymine from deaminated
cytosine. But in RNA the original Uracil and
the deaminated product of cytosine are
undistinguishable and hence incapable of
repair.]
Both DNA and RNA are able to mutate. In
fact, RNA being unstable, mutate at a faster
rate. This is the reason why viruses having
RNA as genome and having shorter life span
mutate and evolve faster.
The DNA and not the RNA forms the
chromosomes. So, only the DNA is able to
express itself in the form of Mendelian
Characters.
Concluding from these reasons it is clear that
while both DNA and RNA can act as the
genetic material, DNA seems to appear a
better candidate molecule for genomic
information storage, especially in higher
organisms.
RNA WORLD
Since, it is clear that RNA as well as DNA are
found as genetic material in organisms, with
RNA genomes being more frequently
observed in primitive species like some
bacteriophages, an immediate logical
question blooms as to whether DNA had
replaced RNA in course of evolution due to
its selective advantages over the latter.
The phrase "The RNA World" was coined by
Walter Gilbert in 1986 on the then recent
observations of the catalytic properties of

various RNAs. In fact, the RNA which was

initially thought to be only a passive
messenger molecule was found in many
active catalytic roles like adapters,
Ribozymes, riboswitches, regulatory
molecules and so on. As the RNA was found
to be associated with such a variety of
functions and essential life processes like
metabolism, translation or splicing, it was
hypothesized logically that this very
biological molecule should have been the
carrier, executor and the maintainer of life,
as it began. Some of the many roles in the
sustenance of life could have been then
distributed over to DNA (which appeared to
be a better candidate for information
storage) and to proteins (which appeared to
be way too better in folding and catalysis) as
evolution progressed. The RNA World refers
to this hypothetical stage in the origin of life
on Earth during which, proteins were not yet
engaged in biochemical reactions and RNA
carried out both the information storage task
of genetic information as well as the full
range of catalytic roles necessary in a very
primitive self-replicating system.
This is the main logic behind the hypothesis.
DNA REPLICATION
The copying mechanism for the molecule is
inherent in its property of complementary
base pairing by which each of the two
strands would separate and act as a
template for the synthesis of new
complementary strands. After the completion
of replication, each DNA molecule would
have one parental and one newly
synthesized strand. This scheme was termed
as semi-conservative DNA replication.
The Experimental Proof for semiconservative replication
Matthew Meselson and Franklin Stahl
performed the following experiment in 1958:

does not initiate randomly at any place in the

DNA but at specific sequences called ori
(which have double H-bonded A-T rich
regions and are hence relatively easier to
separate than triple H-bonded C-G regions).
Such points in the DNA are known as the
origin of replication. While prokaryotes
have generally only one such point,
eukaryotes have hundreds of them.
Initiation and Unwinding

A culture of E.coli bacteria (prokaryote)

grown for several generations on heavy
nitrogen N15 source was introduced on a
medium having only N14 source and the
density of DNA at each subsequent
generation (replication) was studied.
A similar experiment was performed on the
beans, Vicia faba (a eukaryote) by Taylor and
colleagues in 1958 using radioactive
Thymidine.
The experiments proved that the free DNA as
well as DNA packed within chromosomes,
both replicate semi-conservatively.

During initiation, initiator proteins bind to

the stretch of DNA called the replication
origin, thus triggering events that unwind the
DNA double helix into two single-stranded
DNA molecules.
DNA helicases are responsible for breaking
the hydrogen bonds that join
the complementary nucleotide bases to each
other. Because the newly unwound single
strands have a tendency to rejoin, another
group of proteins, the Single-strandbinding proteins, keep the single strands
stable and separated until elongation begins.
A third family of proteins, the
Topoisomerases, which includes Gyrase,
reduces the torsional strain caused by the
unwinding of the double helix.

The Molecular Machinery of Bacterial

DNA Replication
Even while a prokaryotic cell has its genome
built out of a few million base pairs, which is
relatively short in comparison to eukaryotes,
which have billions of base pairs, DNA
replication involves an incredibly
sophisticated, highly coordinated series of
molecular events, even in bacteria. These
events can be divided into four major stages:
initiation, unwinding, primer synthesis and
elongation.
Unwinding and separating the entire length
of DNA is energetically herculean, if not
impossible and so the instantaneous act of
replication occurs only within a small
opening of the DNA helix, referred to as
replication fork. Also the replication fork

The entire process of replication becomes

possible only because of the concerted
activity of a team of many such proteins
which form a multifunctional complex or a
replication machine.

Primer synthesis and Elongation

At the heart of the replication machine is an
enzyme called DNA Polymerase III often
referred to as DNA-dependent DNA
polymerase (since it uses a DNA template to
catalyze the polymerization of
deoxyribonucleotides). This enzyme is very
fast (joins 2000 nucleotides s-1 and is very
efficient (have proof-reading activity). Mg2+
is the cofactor for the enzyme.
Deoxyribonucleoside triphosphates serve
dual purposes here. They are the substrates
as well as the fuel for the reaction (same as
in case of ATP).
But even then, the DNA polymerases, on
their own, can only add deoxyribonucleotides
to the 3'-OH group of an existing chain and
cannot begin synthesis de novo. So, an
enzyme called a DNA Primase (which is
essentially a RNA polymerase) fixes a
temporary primer (short stretches of RNA)
initially, for the polymerase to start working.
Later, these primer fragments are replaced
by DNA Polymerase I and the sugarphosphate backbone stitched up by DNA
ligase.
The DNA-dependent DNA polymerases
catalyze polymerization only in one direction,
i.e. 5'3', by extending the 3OH of the
growing polymer. This directionality is a
consequence of the need for proof-reading
activity.
This directionality also creates some
additional complications at the replicating
fork.
On one strand (the template with polarity
3'5'), the replication is continuous, while
on the other (the template with polarity
5'3'), it is discontinuous. The
discontinuously synthesized (Okazaki)
fragments are later joined by the enzyme
DNA ligase.

*In eukaryotes, the replication of DNA takes

place at S-phase of the cell-cycle.
TRANSCRIPTION and TRANSLATION
Determination of the structure of the DNA
gave unmistakable clues to the fact that the
hereditary information in cells must be
encoded in DNAs sequence of nucleotides.
This code would be the basis for the
production of the molecules which make
biological life possible: the protein and the
RNA. Thus DNA was conferred the title, the
blueprint of life. Replication is the means of
the safe transfer of this coded information to
subsequent generations while transcription
and translation are the 2 steps by which the
cell decodes and uses this information to
make life happen: direct the formation,
development and sustenance of every form
of life, be it a bacterium, a fruit fly, or a
human.
Transcription and translation stands in the
way between the genotype and the
phenotype of a living organism. Since
proteins are the principal constituents of
cells, they determine the structure as well as
the functions of the cell at the immediate
visible level and are in fact the molecular
basis for phenotype. As we know that
proteins are the ultimate form of expression
of the DNA codes, the genetic instructions
carried by the DNA must therefore specify
the amino acid sequences of proteins and
somehow direct their production.
DNA does not direct protein synthesis itself,
but acts rather like a manager, delegating
the various tasks to a team of workers. When

a particular protein is needed by the cell, the

nucleotide sequence of the appropriate
section of the immensely long DNA molecule
in a chromosome is first copied into a more
versatile type of nucleic acid - the RNA
(transcription). These RNA copies of short
segments of the DNA may then be used to
direct the synthesis of the protein
(translation). . In some cases, the RNA
molecule itself is a "finished product" that
serves some important function within the
cell.
All cells, from bacteria to humans, express
their genetic information in this waya
principle so fundamental that it has been
termed the central dogma of molecular
biology.
History: Discovering the Relationship
between DNA and Protein Production
Gene-protein connection
In 1902, Archibald Garrod, recorded
observations of Alkaptonuria patients, whose
urine turned black due to the buildup of a
chemical called Homogentisate. Knowledge
of the biochemical pathway of the
phenylalanine metabolism, made it clear to
him that Homogentisate was one of the
intermediates through which the amino acid
ultimately got degraded into
maleylacetoacetate. This led him to surmise
that the enzyme homogentisate oxidase,
which metabolized homogentisate, must be
defective in his patients.
Correlating with the information that
Alkaptonuria followed a recessive Mendelian
inheritance pattern, Garrod made an even
bolder prediction that a defective gene must
be responsible for the defective enzyme.
Garrod's proposition, attributing a defective
enzyme to a defective gene, was the first
ever to suggest a direct link between genes
and proteins. All that was known at the stage
was that the genetic material (DNA) was
housed in the nucleus within chromosomes.
The proposition led investigators to

subsequently suggest that the nucleus could

also be the site of protein synthesis.
Site of protein synthesis
The exact site of protein synthesis was
confirmed as being the cytoplasm only after
serious investigations involving an alga
called Acetabularia, whose interesting life
cycle posed a stage that created an
opportunity in which the nucleus could be
removed without causing major damage to
the cell. After the removal of the nucleus, the
cells protein production was measured over
time. The unexpected discovery was that the
enucleated alga could still live for months.
Protein production did not stop
instantaneously, pointing out that nucleus
was not the direct site as previously thought.
But, the production ceased within 2 weeks,
indicating that nucleus did in fact have some
role in the long-term production of proteins.
Missing link between DNA and protein
Although Garrod and several other scientists
had demonstrated a clear association
between genes (which were known to be on
chromosomes in the nucleus) and proteins,
the precise nature of this link remained
mysterious for some time. Researchers
wondered whether chromosomes
participated directly in protein production. If
so, one would expect that some DNA would
be found beyond the nucleus, in the
cytoplasm, at least some of the time.
However, no evidence of DNA outside the
nucleus had ever been found. Thus, the
exclusive localization of DNA to the nucleus
could only be linked to protein synthesis in
the cytoplasm if there were some kind of
intermediate messengera substance
"between" the DNA in the nucleus and the
protein production machinery in the
cytoplasm. The early work of Brachet et al
with dyes predicted that another type of
nucleic acid, ribonucleic acid (RNA), might be
the intermediary. Several pieces of evidence
implicated RNA in protein production,
including the following:

RNA is found in both the nucleus and the

cytoplasm.
RNA concentration correlates with protein
production.
Cells that produce large amounts of protein
had cytoplasmic dye- and radiationabsorbing regions indicative of the presence
of nucleic acids and the treatment of such
cells with ribonuclease decreased the cells'
dye- and radiation-absorbing regions.

The process of copying genetic information

from one of the strands of the DNA into RNA
is termed as transcription.
Here also, the principle of complementarity
governs, except that adenosine now forms
base pair with Uracil instead of thymine.
However, unlike in the process of replication,
which once set in, the total DNA of an
organism gets duplicated, in transcription
only a segment of DNA and only one of the
strands is copied into RNA. But, the strand of
DNA that serves as the coding template for
one gene may be non-coding for
other genes within the
same chromosome.This necessitates defining
the boundaries that would demarcate the
region and the strand of DNA that would be
transcribed.
Transcription Unit
A transcription unit in DNA is defined
primarily by the three regions in the DNA:
(i) A Promoter
(ii) The Structural gene
(iii) A Terminator
There is a convention in defining the two
strands of the DNA in the structural gene of a
transcription unit. Since the two strands have
opposite polarity and the DNA-dependent
RNA polymerase also catalyze the

polymerization in only one direction, that is,

5'3' , the strand that has the polarity 3'5'
acts as a template, and is also referred to as
template strand. The other strand which
has the polarity (5'3') and the sequence
same as RNA (except thymine at the place of
uracil), is displaced during transcription.
Strangely, this strand (which does not code
for anything) is referred to as coding
strand. All the reference point while defining
a transcription unit is made with coding
strand.
The promoter and terminator flank the
structural gene in a transcription unit. The
promoter is said to be located towards 5'-end
(upstream) of the structural gene (the
reference is made with respect to the
polarity of coding strand). It is a DNA
sequence that provides binding site for RNA
polymerase, and it is the presence of a
promoter in a transcription unit that also
defines the template and coding strands. By
switching its position with terminator, the
definition of coding and template strands
could be reversed.
The terminator is located towards 3'-end
(downstream) of the coding strand and it
usually defines the end of the process of
transcription. There are additional regulatory
sequences that may be present further
upstream or downstream to the promoter.
Transcription Unit and the Gene
A gene is defined as the functional unit of
inheritance. Though there is no ambiguity
that the genes are located on the DNA, it is
difficult to literally define a gene in terms of
DNA sequence. The DNA sequence coding for
tRNA or rRNA molecule also define a gene.
However since a cistron is defined as a
segment of DNA coding for a polypeptide,
the structural gene in a transcription unit
could be said as monocistronic (mostly in
eukaryotes) or polycistronic (mostly in
bacteria or prokaryotes).

In eukaryotes, the monocistronic structural

genes have interrupted coding sequences
the genes in eukaryotes are split. The coding
sequences or expressed sequences are
defined as exons. While the intervening
sequences which do not appear in the
mature or processed RNA are called introns.
The split-gene arrangement further
complicates the definition of a gene in terms
of a DNA segment.
Inheritance of a character is also affected by
promoter and regulatory sequences of a
structural gene. Hence, sometime the
regulatory sequences are loosely defined as
regulatory genes, even though these
sequences do not code for any RNA or
protein.
Types of RNA and the process of
Transcription
In bacteria, there are three major types of
RNAs: mRNA (messenger RNA), tRNA
(transfer RNA), and rRNA (ribosomal RNA). All
three RNAs are needed to synthesize a
protein in a cell. The mRNA provides the
template, tRNA brings amino acids and reads
the genetic code, and rRNA play structural
and catalytic role during translation.

steps, which are initiation, elongation and

termination.
The RNA polymerase is only capable of
catalyzing the process of elongation. It
associates transiently with initiation-factor
() and termination-factor () to initiate
and terminate the transcription, respectively.
In bacteria, since the mRNA does not require
any processing to become active, and also
since transcription and translation take place
in the same compartment (there is no
separation of cytosol and nucleus in
bacteria), many times the translation can
begin much before the mRNA is fully
transcribed. Consequently, the transcription
and translation can be coupled in bacteria.
In eukaryotes, there are two additional
complexities
(i) There are at least three RNA polymerases
in the nucleus (in addition to the RNA
polymerase found in the organelles). There is
a clear cut division of labor.

The RNA polymerase I transcribes

ribosomal RNA (rRNA)

The RNA polymerase III transcribes tRNA,

one small ribosomal RNA (5srRNA),
and snRNA (small nuclear RNAs)involved in RNA splicing and gene
regulation.

The RNA polymerase II transcribes the

precursor of mRNA, the heterogeneous
nuclear RNA (hnRNA).

There is single DNA-dependent RNA

polymerase that catalyses transcription of all
types of RNA in bacteria.
RNA polymerase binds to promoter and
initiates transcription (Initiation). It uses
nucleoside triphosphates as substrate and
polymerizes in a template depended fashion
following the rule of complementarity. It
somehow also facilitates opening of the helix
and continues elongation. Only a short
stretch of RNA remains bound to the enzyme.
Once the polymerase reaches the terminator
region, the nascent RNA as well as the RNA
polymerase falls off. This results in
termination of transcription.
An intriguing question is that how is the RNA
polymerases able to catalyze all the three

(ii) The second complexity is that the primary

transcripts contain both the exons and the
introns and are non-functional. Hence, it is
subjected to a process called splicing where
the introns are removed and exons are joined
in a defined order. Also the hnRNA undergo
two additional processing called as capping
and tailing. In capping an unusual
nucleotide (methyl guanosine triphosphate)
is added to the 5'-end of hnRNA. In tailing,

adenylate residues (200-300) are added at

3'-end in a template independent manner.
The fully processed hnRNA, now called
mRNA, is transported out of the nucleus for
translation.
The significance of such complexities is now
beginning to be understood. The split-gene
arrangements represent probably an ancient
feature of the genome. The presence of
introns is reminiscent of antiquity, and the
process of splicing represents the dominance
of RNA-world. In recent times, the
understanding of RNA and RNA-dependent
processes in the living system have assumed
more importance.