Bio Informatics

Lecture 2
 Transcription & Translation
• General definition:
• Transcription is synthesis of single stranded RNA from a double
stranded DNA template. Its produces messenger RNA ( mRNA).

• Translation is the 1st stage of protein biosynthesis from RNA. In this

process formation of a polypeptide by using mRNA as a template. It
occurs in ribosomes.
• Transcription and Translation both process are the part of gene
expression.
 Transcription
• Transcription is the synthesis of mRNA from a DNA template which
occurs in 5’-3’ direction.
• During transcription , a DNA sequence is read by an RNA polymerase
which produce a complementary and antiparallel RNA strand.
• Transcription is the first step leading to gene expression.
• The stretch of DNA transcribed into an RNA molecule is called as
transcription unit which encoded at least one gene.
• The result of the transcription is a mRNA which will then be used to
create that protein via the process of translation
 Transcription – Eukaryotes v/s Prokaryotes
• Prokaryotic transcription occurring in cytoplasm alongside translation
and eukaryotic transcription occurring only in the nucleus where it is
separated from the cytoplasm by the nuclear membrane.
• Eukaryotic DNA not currently used in stored as heterochromatin
around histones to from nucleosomes and must be unwound as
euchromatin to be transcribed.
 Transcription - Stages
• There are three stages involved in transcription :-
• A. INITIATION
• B. ELONGATION
• C. TERMINATION
 Initiation - Transcription
• RNA polymerase binds to specific DNA region and initiate
transcription called as promoter site.
• RNA polymerase is the enzyme responsible for transcription.
• After polymerase is bound to the promoter DNA, the two DNA strands
unwind and the enzyme starts transcribing the template strand.
• The position of the first synthesized base of the RNA is called the start
site.
 Elongation - Transcription
• RNA polymerase moves along DNA template and sequentially
synthesizes the RNA chain.
• DNA is unwinding ahead of the moving polymerase and the helix is
reformed behind it. It unwinds 10-20 DNA bases at a time.
• RNA polymerase add nucleotides in the 5’-3’ direction.
• The new section of RNA ‘peels away’ as the double helix reforms.
 Termination - Transcription
• Transcription stops when RNA polymerase reaches a section of DNA
called the terminator.
• Terminator sequence = AAUAAA.
• Next, the RNA strand is released and RNA polymerase dissociates
from the DNA.
• The RNA strand will go through more processing.
 Pre mRNA to mRNA - Processing
• The original transcript from the DNA is called as pre-m RNA.
• It contains transcript of both intron and exons.
• Intron:- it is non-coding sections of nucleic acid found between
coding regions.
• Exons:-coding regions of nucleic acids
• Pre-mRNA never leaves the cell’s nucleus.
• The introns are excised and exons are joined together to form mRNA.
• The introns are removed by a process called splicing to produce
messenger RNA (mRNA)
 Translation
• Translation is a process in which the formation of polypeptide
(PROTEIN) by decoding mRNA produced in transcription.
• It occurs in ribosome which are present in cytoplasm.
• It begins after mRNA enters in cytoplasm.
• It uses tRNA as the interpreter of mRNA.
 Ribosomes – Tiny Protein Factories
• Ribosomes are the structures where polypeptides (proteins) are built.
They are made up of protein and RNA (ribosomal RNA, or rRNA).
• Each ribosome has two subunits, a large one and a small one, which
come together around an mRNA—kind of like the two halves of a
hamburger bun coming together around the patty.
• The ribosome provides a set of handy slots where tRNAs can find their
matching codons on the mRNA template and deliver their amino acids.
• These slots are called the A, P, and E sites.
• Not only that, but the ribosome also acts as an enzyme, catalyzing the
chemical reaction that links amino acids together to make a chain.
 Translation - Stages
• Translation proceed in four stages:-
• 1. INITIATION
• 2. ELONGATION
• 3. TRANSLOCATION
• 4. TERMINATION
 Initiation - Translation
• The initiation stage of translation brings together mRNA, tRNA bearing the first amino
acid of the polypeptide, and two subunits of a ribosome.
• The components involved are the large and small subunits of ribosome, mRNA, initiator
tRNA in its charged form and three factors (IF1.IF2,IF3) and GTP.
• The tRNA has a amino acid linked to it is term as Charged tRNA.
• IF1 and IF2 bind to free 30S subunit.
• IF3 complexed with GTP then bind to the small subunit. It will assist the charged initiator
tRNA to bind.
• The assembled ribosome has 2 tRNA binding sites. These are called A-site (acceptor) for
aminoacyl and P-site(donor)for polypeptide.
• The A-site is where incoming aminoacyl-tRNA moleules bind and P-site where the
growing polypeptide chain usually found.
 Initiation – Contd.
• One major outcome of initiation is the placement of initiator tRNA in
the P-site.
• Start codon :- AUG
• Start anticodon :- UAC
• The small ribosomal subunit attaches to 5’ end of mRNA.
 Elongation - Translation
• In this amino acid are added one by one to the first amino acid called
as amino acid delivery.
• In codon recognition , mRNA codon in the A site forms hydrogen bond
with the tRNA anticodon.
• In peptide bond formation, the ribosome catalyzes the formation of
the peptide bond between the amino acids. The polypeptide
extending from the Psite moves to A-site to attach to the new Amino
Acid.
• In elongation process three elongation factors (EF-T4,EF-T5,EF-G)
which will bind with GTP or GDP.
 Translocation - Translation
• The t-RNA with the polypeptide chain in the A site is trans located to
the P site. tRNA at the P site moves to the E site and leaves the
ribosome.
• The ribosome moves down the mRNA in the 5’-3’ direction.
 Transfer RNAs - tRNAs
• ransfer RNAs, or tRNAs, are molecular "bridges" that connect mRNA
codons to the amino acids they encode.
• One end of each tRNA has a sequence of three nucleotides called an
anticodon, which can bind to specific mRNA codons.
• The other end of the tRNA carries the amino acid specified by the
codons.
• There are many different types of tRNAs.
• Each type reads one or a few codons and brings the right amino acid
matching those codons.
Transfer RNAs - tRNAs
 Termination - Translation
• Protein factors called release factors interact with the stop codons
and cause release of the completed polypeptide chain.
• Stop codon – UAA,UAG,UGA.
• RF1 recognizes the codons UAA and UAG. RF2 recognizes UAA and
UGA. RF3 helps either RF1 and RF2 to carry out the reaction.
 Codons - The Genetic code
• During translation, a cell “reads” the information in a messenger RNA
(mRNA) and uses it to build a protein.
• Actually, to be a little more technical, an mRNA doesn’t always
encode—provide instructions for—a whole protein.
• Instead, what we can confidently say is that it always encodes a
polypeptide, or chain of amino acids.
• In an mRNA, the instructions for building a polypeptide are RNA
nucleotides (As, Us, Cs, and Gs) read in groups of three. These groups
of three are called codons.
 Codons - The Genetic code
• There are 61 codons for amino acids, and each of them is "read" to
specify a certain amino acid out of the 20 commonly found in proteins.
• One codon, AUG, specifies the amino acid methionine and also acts as a
start codon to signal the start of protein construction.
• There are three more codons that do not specify amino acids.
• These stop codons, UAA, UAG, and UGA, tell the cell when a
polypeptide is complete.
• All together, this collection of codon-amino acid relationships is called
the genetic code, because it lets cells “decode” an mRNA into a chain of
amino acids.
Codons - The Genetic code
Codon Table - The Genetic code
 Useful Links
• https://courses.lumenlearning.com/boundless-biology/chapter/eukar
yotic-transcription
/

• https
://courses.lumenlearning.com/boundless-biology/chapter/ribosomes-
and-protein-synthesis
/