Protein Synthesis-
the ”Stuff of Life“
Translation-
Making the Protein
Manish Jain
Protein Synthesis
Protein biosynthesis (or protein synthesis) is a core
biological process, occurring inside cells, balancing
the loss of cellular proteins (via degradation or export)
through the production of new proteins.
Protein synthesis can be divided broadly into two
phases - transcription and translation.
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Key Terms
RNA (ribo- Single-stranded nucleic acid that carries out the
nucleic acid) instructions coded in DNA
Central dogma The process by which the information in genes flows
of biology into proteins: DNA — RNA = protein
Polypeptide A chain of amino acids
Codon A sequence of three nucleotides that corresponds
with a specific amino acid or start/stop signal during
translation
Transcription Process during which a DNA sequence of a gene is
copied to make an RNA molecule
Translation Process during which an mRNA molecule is used to
assemble amino acids into polypeptide chains
Mutation A change in a genetic sequence
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DNA, RNA AND NUCLEOTIDE
• All DNAs and most RNAs are long, unbranched polymers of nucleotides.
• A nucleotide consists of a phosphorylated pentose linked to
an organic base, either a purine (AG) or a pyrimidine (CT).
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Protein Synthesis
• Protein synthesis consists of two processes —
transcription and translation
• Transcription occurs in the nucleus. Transcription is the
process of copying a segment of DNA into RNA
• During transcription, DNA is used as a template to make
a molecule of messenger RNA (mRNA).
• During translation, the genetic code in mRNA is read
and used to make a protein. These two processes are
summed up by the central dogma of molecular
biology: DNA → RNA → Protein.
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Protein Synthesis
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Protein Synthesis
• The segments of DNA transcribed into RNA molecules
that can encode proteins are said to produce messenger
RNA (mRNA). Other segments of DNA are copied into
RNA molecules called non-coding RNAs (ncRNAs) such
as microRNA, transfer RNA (tRNA).
• mRNA comprises only 1–3% of total RNA samples
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Protein Synthesis
• In eukaryotes, this mRNA molecule is known as pre-
mRNA as it undergoes post-transcriptional modifications
in the nucleus to produce a mature mRNA molecule.
• During translation, the genetic code in mRNA is read
and used to make a protein. These two processes are
summed up by the central dogma of molecular
biology: DNA → RNA → Protein.
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Protein Synthesis
• A DNA transcription unit encoding for a protein may
contain both a coding sequence, which will be
translated into the protein, and regulatory sequences,
which direct and regulate the synthesis of that protein.
• The regulatory sequence before ("upstream" from) the
coding sequence is called the five prime untranslated
region (5'UTR); the sequence after ("downstream" from)
the coding sequence is called the three prime
untranslated region (3'UTR).
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Protein Synthesis
• Place - mRNA (nucleus) → ribosome (cytoplasm), where
translation occurs.
The process of
transcription
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Transcription
• The production of a RNA copy from a DNA strand is called
transcription
• Transcription is the first part of the central dogma of
molecular biology: DNA → RNA.
• It is the transfer of genetic instructions in DNA to mRNA.
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Transcription
• During transcription, a strand of mRNA is made to
complement a strand of DNA.
The strands of the double helix must separate to allow the
internal edges of the bases to pair with the bases of the
nucleotides being polymerized into new complementary
polynucleotide chains. 12
Transcription
• an enzyme known as a helicase, binds to DNA and
breaks a hydrogen bond in one strand.
phosphodiester bond
The 3' carbon of one sugar is bonded to the 5' phosphate of the
adjacent sugar. 13
Transcription
• Such a one-strand break in DNA is called a nick, leading
to loss of supercoils.
• The same enzyme joins (ligates) the two ends of the
broken strand 14
Transcription
• Transcription uses one of the two exposed DNA strands
as a template; this strand is called the template strand.
• The RNA product is complementary to the template
strand and is almost identical to the other DNA strand,
called the nontemplate (or coding) strand.
• One important
difference: in RNA,
all of the T
nucleotides are
replaced with U
nucleotides
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Steps of Transcription
3 steps:
•Initiation
•Elongation
•Termination.
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Steps of Transcription- Initiation
1.Initiation is the beginning of
transcription. It occurs when
the enzyme RNA polymerase
binds together with one or more
general transcription factors to a
region of a gene called
the promoter.
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Promoter
1.Recognition sites contain specific sequences (palindromic)
of nucleotides (4-8 base pairs in length)
2.Sequence consisting of two identical or highly similar
inverted repeats which are adjacent to one another or
separated by a spacer region
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RNA polymerases
Three types of RNA polymerases, each of which needs a
special DNA sequence (promoter) and a set of DNA-
binding proteins (transcription factors).
1.RNA polymerase I- transcription of ribosomal RNA (rRNA)
genes.
2.RNA polymerase II- transcribes all protein-coding genes
3.RNA polymerase III- transcribes transfer RNA (tRNA)
genes,
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Steps of Transcription- Initiation
• Initiation- RNA polymerase binds to promoter. This signals
the DNA to unwind (approximately 14 base pairs) so
the enzyme can “read” the bases in one of the DNA strands.
• The DNA uncoils
(transcription bubble)
and provides the pattern
for the formation of a
single strand of mRNA.
Steps of Transcription- Initiation
• RNA polymerase, assisted by one or more general
transcription factors, then selects a transcription start site in
the transcription bubble,
• The enzyme makes a
strand of mRNA by
catalyzing phosphodiester
linkage with two initial
rNTP.
• Transcription initiation is
regulated by additional
proteins, known as
activators and repressors,
which modulate
formation and function of
the transcription initiation
complex
Steps of Transcription- Elongation
2. Elongation One strand of the DNA (template), is used for
RNA synthesis.
• Is the addition of
nucleotides to the
mRNA strand.
• RNA polymerase
moves through the
template strand in
the direction of 3'
→ 5’
• Behind the moving RNA polymerase, the two strands of
DNA rejoin. 22
Steps of Transcription- Elongation
• Multiple RNA polymerases can bind on a single DNA
template → multiple rounds of transcription → many
mRNA molecules can be rapidly produced from a
single copy of a gene
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Steps of Transcription- Termination
RNA polymerase will keep
transcribing until it gets signals
to stop.
The process of ending
transcription is
called termination.
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Steps of Transcription- Termination
Before STOP codon, RNA polymerase transcribes a
sequence of DNA known as a terminator.
Terminator is a sequence in the newly synthesized mRNA
that is rich in the nucleotides, guanine and cytosine,
followed by a short stretch that is rich in the nucleotide
uracil
STOP CODON 25
Steps of Transcription- Termination
The stretch of cytosines and guanines in the mRNA can
anneal, creating a double-stranded RNA structure referred
to as a hairpin loop
Once the hairpin is formed in
the nascent mRNA, RNA
polymerase will dissociate
from the DNA and release
the nascent mRNA.
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Translation
• Translation is the second part of the central
dogma of molecular biology: RNA → Protein. It is
the process in which the genetic code in mRNA is
read to make a protein.
• The site of RNA synthesis (nucleus) is separated
from the site of translation (cytoplasm).
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Translation
• Export of RNAs requires association
with specific proteins known as
exportins. Specific exportin are
responsible for the export of a
given RNA type.
• Exportins bind -signaling
sequences called nuclear export
signals (NES) and RanGTP complex
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Translation
• Messenger RNA (mRNA) is translated into protein by the joint
action of transfer RNA (tRNA) and the ribosome, which is
composed of numerous proteins and two major ribosomal RNA
(rRNA) molecules.
• Ribosomes -2 subunits (small
and a large).
• There are three sites where
the tRNA attaches.
•A site- tRNA arrives with the
amino acid.
•P site- tRNA binding site.
•E site -tRNA exits without an
amino acid.
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Translation
• mRNA transcript possess start codon (AUG, specific
sequence of nucleotides) which is responsible for the
recruitment of a ribosome during the initiation of
translation
• Ribosome attaches to mRNA through a ribosome
binding site (RBS, sequence of nucleotides upstream of the
start codon of mRNA
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Translation
• With the help of rRNA, bonds form between the amino
acids as they are brought one by one to the ribosome,
creating a polypeptide chain. The chain of amino acids
keeps growing until a stop codon is reached.
• The ribosome reads the
sequence of codons in
mRNA,
• Molecules of tRNA
bring amino acids to the
ribosome in the correct
sequence.
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Translation
• Transfer RNA (tRNA) is the key to deciphering the code
words in mRNA.
• The tRNA is a small
coiled molecule that
accepts an amino acid on
one end and matches up
to a specific three-
base portion of the
mRNA on the other end.
• Each type of amino acid has its own type of tRNA, which binds it
and carries it to the growing end of a polypeptide chain.
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Translation
• The tRNA interacts with the mRNA so as to put the amino
acid in the proper sequence for the developing protein. After
adding the amino acid to the sequence, the tRNA is then
cleaved from the amino acid and recycled for further use in
the process.
• The correct tRNA with its
attached amino acid is
selected at each step
because each specific
tRNA molecule contains a
three-base sequence that
can base-pair with
its complementary code
word in the mRNA 33
Translation
• The process of protein synthesis is fairly fast.
• Amino acids are added to the growing peptide chain at a
rate of about 3-5 amino acids per second.
• A small protein (100-200 amino acids) can be produced in
a minute or less.
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What Happens Next?
• After a polypeptide chain is synthesized, it may
undergo additional processes.
• Folding
• It may also bind with other polypeptides or with
different types of molecules, such
as lipids or carbohydrates.
• Many proteins travel to the Golgi apparatus within
the cytoplasm to be modified for the specific job.
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Summary
• Protein synthesis is the process in which cells make proteins.
It occurs in two stages: transcription and translation.
• Transcription is the transfer of genetic instructions in DNA to
mRNA in the nucleus. It includes three steps: initiation,
elongation, and termination. After the mRNA is processed, it
carries the instructions to a ribosome in the cytoplasm.
• Translation occurs at the ribosome, which consists of rRNA
and proteins. In translation, the instructions in mRNA are
read, and tRNA brings the correct sequence of amino acids
to the ribosome. Then, rRNA helps bonds form between the
amino acids, producing a polypeptide chain.
• After a polypeptide chain is synthesized, it may undergo
additional processing to form the finished protein.
• Mutations
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