(L-6) - Molecular Basis of Inheritance - Jan 25, 2020

Molecular Basis of Inheritance
DNA Transcription
LECTURE 6
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DNA Transcription
LECTURE 6
Protein Synthesis
Protein Synthesis
➢ Gene expression for protein synthesis is completed by Central Dogma which was
given by Francis Crick.
➢ The process of copying information from one strand of DNA into mRNA is called
Transcription.
➢ Formation of polypeptide using the information from mRNA is called Translation.
Protein Synthesis
Protein Synthesis
Transcription
➢ It is the process of copying genetic information from template strand of DNA into
RNA.
➢ It is mediated by RNA polymerase.
➢ Transcription takes place in the nucleus of eukaryotic cells.
Protein Synthesis
Transcription
➢ Strand of DNA with polarity 3' → 5' that directs synthesis of RNA, is called template
strand or antisense strand.
➢ Other strand of DNA with polarity 5' → 3' is complementary to the template strand is
called coding strand or sense strand.
Protein Synthesis
Transcription
➢ Genetic code present in this strand is similar to genetic code (based on mRNA)
except that uracil (U) is replaced by thymine (T).
➢ It means the coding strand of DNA has the same sequence of nitrogenous bases as
the RNA transcript except that in RNA T (thymine) is substituted by U (uracil).
➢ It has the same polarity as the RNA transcript.
Protein Synthesis
Transcription Unit
➢ It is defined as the segment of DNA between the sites of initiation and termination of
transcription by RNA polymerase.
➢ More than one gene may reside in a transcription unit.
Protein Synthesis
Transcription Unit
➢ Transcription unit in DNA molecule comprises three regions.
Promoter
Structural Gene
Terminator
Transcription Unit
Promoter
➢ It is a region on DNA molecule to which an RNA polymerase binds and initiates

transcription.
➢ Promoter is located upstream (to the left) of the structural gene, i.e., towards 5' end
of coding strand, 3' end of template strand.
➢ Location of promoter gene and terminator gene is always mentioned with reference
to coding strand.
Transcription Unit
Promoter
➢ It has different parts for attachment to various transcription factors.

➢ In most eukaryotes, the promoter has an AT rich region called TATA box.
Transcription Unit
Promoter
➢ The area has a groove to which specific protein components can combine.
➢ TATA box is also called Pribnow box (after its discoverer Pribnow) in prokaryotes
and Hogness box (after its discoverer Hogness) in eukaryotes.
➢ Besides a promoter, eukaryotes also require an enhancer.
TATA Box
Pribnow Box Hogness Box

Transcription Unit
Terminator
➢ This region on DNA molecule is present downstream of the structural gene, i.e.,
towards 3' end of coding strand, 5' end of template strand.
➢ Terminator usually defines the end of the process of transcription.
Transcription Unit
Structural Gene
➢ Area of template strand that is involved in transcription or formation of RNA.
Structural Gene
Monocistronic Polycistronic
Transcription Unit
Structural Gene
Monocistronic Polycistronic
➢ It is made up of only one cistron. ➢ It is made up of many cistrons.

➢ It carries information for synthesis of ➢ It carries information for synthesis of
one polypeptide chain. more than one polypeptide chains.
➢ It is mostly found in eukaryotes. ➢ They are mostly found in prokaryotes.
Protein Synthesis
Transcription Unit and the Gene
➢ A gene is defined as the functional unit of inheritance.

➢ Gene are located on DNA and it is difficult to literally define a gene in terms of DNA
sequence.
➢ DNA sequence coding for tRNA or rRNA molecule also defines a gene.
➢ Cistron is defined as a functional unit of gene, it is a segment of DNA coding for a
polypeptide.
Protein Synthesis
➢ The structural gene in a transcription unit is monocistronic (mostly in eukaryotes)

and polycistronic (mostly in prokaryotes or bacteria).
➢ Monocistronic gene synthesises one type of polypeptide or protein.
➢ Polycistronic gene synthesises different proteins or polypeptides.
Protein Synthesis
➢ The monocistronic structural genes have interrupted coding sequences, i.e., the
genes in eukaryotes are split.
➢ The coding sequences or expressed sequences are defined as exons which appear in
mature or processed RNA.
Protein Synthesis
➢ Exons are interrupted by introns.

➢ Introns are intervening sequences that do not appear in mature or processed RNA.
➢ Split-gene arrangement further complicates the definition of a gene in terms of a
DNA segment.
Protein Synthesis
RNA Synthesis
➢ The mechanism of RNA synthesis was worked out in the late 1950s by Horwitz,
Weiss and Stevens, by in vitro experiments.
➢ No primer is needed for RNA synthesis.
Materials Required for Transcription
Enzyme RNA Polymerase
DNA template
Four types of ribonucleotides triphosphates (ATP, CTP, GTP and UTP)
Divalent metal ions Mg2+ or Mn2+ as a cofactor.

Materials Required for Transcription
RNA Polymerase
Prokaryotes Eukaryotes
➢ There is single RNA polymerase in ➢ Eukaryotes have three types of RNA

prokaryotes which synthesizes all polymerases
types of RNAs. ○ RNA Polymerase I
○ RNA Polymerase II
○ RNA Polymerase III.
RNA Polymerase in Eukaryotes
RNA Polymerase I Synthesizes rRNA (28 S, 18 S) in nucleolus
RNA Polymerase II Synthesizes mRNA and hnRNA
RNA Polymerase III Synthesizes tRNA , 5 srRNA, Sn RNA in nucleolus

Protein Synthesis
RNA Polymerase
➢ It is a large complex consisting of 4 subunits which make the core enzyme and 1 σ
(Sigma) subunit and ρ (Rho) subunit.
RNA Polymerase
Core Enzyme Sigma Subunit Rho Subunit
It has 4 subunits. Recognises Helps in

promoter and termination of
initiates synthesis. transcription.
RNA Polymerase
Core Enzyme
2 ∝ Subunits These bind to regulatory proteins
1 β Subunit It binds to DNA template
1 β’ Subunit It binds to RNA nucleotides

Protein Synthesis
Process of Transcription
Protein Synthesis
Process of Transcription
In Prokaryotes In Eukaryotes
Protein Synthesis
Transcription in Prokaryotes
➢ It occurs in cytoplasm with the help of DNA-dependent RNA polymerase that

catalyses transcription of all the three types of RNAs (mRNA, tRNA and rRNA).
➢ All three RNAs are needed to synthesize a protein in a cell.
Protein Synthesis
➢ In prokaryotes the structural genes are polycistronic and continuous.

➢ The process of transcription is completed in following steps:
Initiation
Elongation
Termination
Protein Synthesis
Initiation
➢ RNA polymerase reaches the promoter region and binds to it.

➢ It has a sigma (σ) factor (also called initiation factor).
➢ The enzyme recognizes the promoter by its sigma factor.
Initiation
➢ RNA polymerase initiates transcription.

➢ It uses nucleoside triphosphates as substrate and polymerases in a template
depended fashion following the rule of complementarity.
➢ It also facilitates opening of helix.
Elongation
➢ RNA polymerase in the presence of energy and Mg2+ can catalyse the process of
elongation.
Elongation
➢ As the enzyme moves along the DNA template, RNA chain becomes longer.
➢ Synthesis of RNA continues till the enzyme reaches the terminator.
Termination
➢ When RNA polymerase reaches terminator region a specific chain terminating

protein called rho (ρ) factor (also called termination factor) stops the synthesis of
RNA chain.
➢ It separates RNA polymerase as well as the newly formed RNA strand.
➢ RNA and RNA polymerase fall off and it results in termination of transcription.
Special Features
a. mRNA does not require any processing to become active.

b. Transcription and translation take place in the same compartment as there is no
separation of cytosol and nucleus.
c. Many times the translation can begin much before the mRNA is fully transcribed.
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Molecular Basis of Inheritance

Dr. Sachin Kapur

20+ years Teaching Experience

➢ Transcription unit in DNA molecule comprises three regions.

➢ It is a region on DNA molecule to which an RNA polymerase binds and initiates

➢ It has different parts for attachment to various transcription factors.

Pribnow Box Hogness Box

➢ Area of template strand that is involved in transcription or formation of RNA.

➢ It is made up of only one cistron. ➢ It is made up of many cistrons.

Transcription Unit and the Gene

➢ A gene is defined as the functional unit of inheritance.

Transcription Unit and the Gene

➢ The structural gene in a transcription unit is monocistronic (mostly in eukaryotes)

Transcription Unit and the Gene

Transcription Unit and the Gene

➢ Exons are interrupted by introns.

Enzyme RNA Polymerase

Four types of ribonucleotides triphosphates (ATP, CTP, GTP and UTP)

Divalent metal ions Mg2+ or Mn2+ as a cofactor.

➢ There is single RNA polymerase in ➢ Eukaryotes have three types of RNA

RNA Polymerase I Synthesizes rRNA (28 S, 18 S) in nucleolus

RNA Polymerase II Synthesizes mRNA and hnRNA

RNA Polymerase III Synthesizes tRNA , 5 srRNA, Sn RNA in nucleolus

Core Enzyme Sigma Subunit Rho Subunit

It has 4 subunits. Recognises Helps in

2 ∝ Subunits These bind to regulatory proteins

1 β Subunit It binds to DNA template

1 β’ Subunit It binds to RNA nucleotides

➢ It occurs in cytoplasm with the help of DNA-dependent RNA polymerase that

➢ In prokaryotes the structural genes are polycistronic and continuous.

➢ RNA polymerase reaches the promoter region and binds to it.

➢ RNA polymerase initiates transcription.

➢ When RNA polymerase reaches terminator region a specific chain terminating

a. mRNA does not require any processing to become active.

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