NE

DA
IV
IM

BIOSINTESIS PROTEIN DAN

ASAM NUKLEAT

23/11/21
Prof. Dr. Ramlan Silaban, M.Si.
Prof. Ramlan Silaban 1
 Ingat kembali :
1. Genotif dan Fenotif
2. Informasi genetik, kode genetik
3. Biosintesis asam-asam amino
4. Biosintesis basa-basa purin dan
pirimidin
5. Biosintesis protein terjadi dalam ribosom
6. Adany 3 jenis RNA (mRNA. tRNA,
rRNA)
7. Human genome
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 The genetic expression :
(by Crik, 1952)

Interrelation of DNA, RNA and Protein

DNA RNA PROTEIN

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 Genetic expression process
 Replication
(DNA biosynthesis)
Tuliskan dalam
 Transcription Buku Catatanmu,
apa arti istilah
(RNA biosynthesis) tersebut, cari
 Translation literaturnya

(Protein biosynthesis)

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 DNA
 Deoxyribonucleic
acid is a coiled
double helix
carrying hereditary
information of the
cell

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 DNA
 Contains the
instructions for
making proteins from
20 different amino
acids
 Appears as chromatin
when cell not dividing

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 DNA
 Structure discovered
by Watson & Crick in
1953
 Sides made of
pentose (5-sided)
sugars attached to
phosphate groups
by phosphodiester
bonds
 Pentose sugar called
Deoxyribose
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 DNA
 Steps or rungs of DNA
made of 4 nitrogen-
containing bases held
together by weak
hydrogen bonds
 Purines (double carbon-
nitrogen rings) include
adenine (A) and guanine
(G)
 Pyrimidines (single
carbon-nitrogen rings)
include thymine (T) and
cytosine (C)

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 DNA
 Base pairing means a purine
bonds to a pyrimidine
(Example: A --- T and C ---
G)
 Coiled, double stranded
molecule known as double
helix
 Make up chromosomes in the
nucleus
 Subunits of DNA called
nucleotides
 Nucleotides contain a
phosphate, a Deoxyribose
sugar, and one nitrogen
base (A,T,C, or G)
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 DNA
 Free nucleotides also
exist in nucleus
 Most DNA is coiled or
twisted to the right
 Left twisted DNA is
called southpaw or Z-
DNA
 Hot spots which can
result in mutations occur
where right & left twisted
DNA meet
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 Types of Gene
 Structural Gene (coding of properties)
 Regulatory Gene

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 Regulatory Gene
1. Initiating (i)
2. Operating (o)
3. Promoting (p)

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 DNA Replication
 Process by which DNA makes
a copy of itself
 Occurs during S phase of
interphase before cell division
 Extremely rapid and accurate
(only 1 in a billion are
incorrectly paired)
 Requires many enzymes &
ATP (energy)
 Begins at special sites along
DNA called origins of
replication where 2 strands
open & separate making  a
replication fork

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 DNA Replication
 Nucleotides added &
new strand forms at
replication forks
 DNA helicase
(enzyme) uncoils &
breaks the weak
hydrogen bonds
between
complementary bases
(strands separate)
  DNA polymerase
adds new nucleotides
to the exposed bases

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953 Leh
 DNA Replication
 DNA ligase helps join
segments together

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 DNA Replication
 DNA polymerase
proofreads the new
DNA checking for
errors & repairing
them; called excision
repair
 Helicase recoils the
two, new identical
DNA molecules

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 RNA
 Ribonucleic acid
 Single stranded
molecule  

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 RNA
 Found in nucleus & cytoplasm
 Contains ribose sugar
 Contains the nitrogen base uracil (U)
instead of thymine so A pairs with U
 Base pairings are A-U and C-G
 Three types of RNA exist (mRNA, TRNA,
& rRNA)

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 Transcription
 Transcription is the process by which an mRNA
template, encoding the sequence of the protein in the
form of a trinucleotide code, is transcribed from the
genome to provide a template for translation.
Transcription copies the template from one strand of the
DNA double helix, called the template strand.
 Transcription can be divided into 3 stages: Initiation,
Elongation and Termination, each regulated by a large
number of proteins such as transcription factors and
coactivators that ensure the correct gene is transcribed
in response to appropriate signals.
 The DNA strand is read in the 3' to 5' direction and the
mRNA is transcribed in the 5' to 3' direction by the RNA
polyermase
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 Initiation in eukaryotes
 The initiation of protein translation in eukaryotes is
similar to that of prokaryotes with some modifications.
 A complex of proteins will connect the 5'cap and 3'PolyA
tail, and this complex will recruit the ribosome subunits.
 There is no Shine-Dalgarno sequence in eukaryotes.
Instead, the ribosome scans along the mRNA for the first
methionine codon. Similarly, there is no N-
formylmethionine in eukaryotic cells.
 Shine-Dalgarno sequence, proposed by Australian scientists John
Shine and Lynn Dalgarno,is a ribosomal binding site generally located
6-7 nucleotides upstream of the start codon AUG. The Shine-Dalgarno
sequence exists only in prokaryotes; the six-base consensus sequence
is AGGAGG

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 Elongation
 Elongation of protein biosynthesis is fairly similar between prokaryotes and
Elongation of protein biosynthesis is fairly similar between prokaryotes and
eukaryotes. The following is a description of elongation in prokaryotes.
 Elongation proceeds after initiation with the binding of an aminoacyl-tRNA to the A
site, which is the next codon in the mRNA. The aminoacyl-tRNA is brought to the
ribosome through a series of interactions with EF-Tu (an elongation factor) set of
proteins that facilitate the events . This step involves the hydrolysis of GTP: EF-Tu-
GTP → EF-Tu-GDP (The hydrolyzed GDP is switched for GTP through another
series of reactions with EF-Ts.)
 The next aminoacyl-tRNA binds to the codon, and the C-terminus of the F-Met
undergoes nucleophilic attack by the N-terminus of the second amino acid. The F-
Met is now connected to the second amino acid through a peptide bond.
 The first tRNA (for F-Met) is now uncharged. The entire ribosome complex moves
along the mRNA through the action of another elongation factor (EF-G) and the
hydrolysis of GTP → GDP.
 The first tRNA is now in the E site and comes off from the ribosome, while the
second tRNA, with the nascent peptide chain, is in the P site. Step 1-4 will repeat as
successive amino acids are added.
 f-Met is always the first amino acid in a polypeptide chain
 N-terminus (also known as the amino-terminus, NH2-terminus, N-terminal end or amine-terminus)
refers to the end of a protein or polypeptide terminated by an amino acid with a free amine group (-NH2).
 The C-terminus (also known as the carboxyl-terminus,
carboxyl-terminus, carboxy-terminus,
carboxy-terminus, C-terminal end,
end, or COOH-
terminus) of a protein or polypeptide is the end of the amino acid chain terminated by a free carboxyl group
(-COOH)
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 Termination
 Termination of protein biosynthesis occurs when
the ribosome comes across a stop codon, for
which there is no tRNA. At this point, protein
biosynthesis halts and one of three release
factors will bind to the stop codon. (Note: In
eukaryotes, there is only one release factor that
will bind to all three stop codons.) This induces a
nucleophilic attack of the C-terminus of the
nascent peptide by water - this hydrolysis
releases the peptide from the ribosome. The
ribosome, release factor, and uncharged tRNA
then dissociate and translation is complete.

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 Transcription
 Transcription starts with an enzyme called polymerase
copying the DNA sequence to a similar molecule called
messenger RNA (mRNA). It replaces T with U (Uracil), a
helper base, making it clear that the mRNA is a copy.
The bases (A, T, G, C) on one strand of the DNA specify
the order of bases on the new strand of mRNA (A, U, G,
C). The DNA stays inside the nucleus, but the mRNA
travels out into the cytoplasm.
 http://www.stolaf.edu/people/giannini/flashanima
t/molgenetics/transcription.swf
 http://vcell.ndsu.nodak.edu/animations/transcription/movi
e.htm

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 Steps in Transcription
 DNA helicase (enzyme) uncoils the DNA molecule
 RNA polymerase (enzyme) binds to a region of DNA
called the promoter which has the start codon TAC to
code for the amino acid methionine
 Promoters mark the beginning of a DNA chain in
prokaryotes, but mark the beginning of 1 to several
related genes in eukaryotes
 The 2 DNA strands separate, but only one will serve as
the template & be copied
 Free nucleotides are joined to the template by RNA
polymerase in the 5’ to 3’ direction to form the mRNA
strand
 mRNA sequence is built until the enzyme reaches an area
on DNA called the termination signal
 RNA polymerase breaks loose from DNA and the newly
made mRNA
 Eukaryotic mRNA is modified (unneeded sections
snipped out by enzymes & rejoined) before leaving the
nucleus through nuclear pores, but prokaryotic RNA
isn’t
 All 3 types of RNA called transcripts are produced by this
method

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 Steps in Translation
 tRNA’s break loose
from amino acid,
leave the ribosome, &
return to cytoplasm to
pick up another amino
acid
 Animation of
translation
 http://sps.k12.ar.us/m
assengale/ANIMPRO
T.htm
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  mRNA
 Messenger RNA
 Single, uncoiled, straight strand of nucleic acid
 Found in the nucleus & cytoplasm
 Copies DNA’s instructions & carries them to the
ribosomes where proteins can be made
 mRNA’s base sequence is translated into the
amino acid sequence of a protein
 Three consecutive bases on mRNA called a
codon (e.g. UAA, CGC, AGU)
 Reusable
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 tRNA
 Transfer RNA
 Single stranded molecule containing
80 nucleotides in the shape of a
cloverleaf
 Carries amino acids in the cytoplasm
to ribosomes for protein assembly
 Three bases on tRNA that are
complementary to a codon on mRNA
are called anticodons (e.g. codon-
UUA; anticodon- AAU)
 Amino Acid attachment site across
from anticodon site on tRNA
 Enters a ribosome & reads mRNA
codons and links together correct
sequence of amino acids to make a
protein
 Reusable  

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 rRNA
 Ribosomal RNA
 Globular shape
 Helps make up the structure of
the ribosomes  
 rRNA & protein make up the
large & small subunits of
ribosomes
 Ribosomes are the site of
translation (making
polypeptides)
 Aids in moving ribosomes
along the mRNA strand as
amino acids are linked
together to make a prote

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 Proteins
 Organic molecules (macromolecules)
made by cells
 Make up a large part of your body
 Used for growth, repair, enzymes, etc.
 Composed of long chains of small units
called amino acids bonded together by
peptide bonds
 Twenty amino acids exist
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 Protein Synthesis
 Consists of 2 parts --- Transcription &
Translation
 Begins in the nucleus with mRNA copying
DNA’s instructions for proteins
(transcription)
 Completed in the cytoplasm when tRNA
enters ribosomes to read mRNA codons
and link together amino acids
(translation)
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 What is protein synthesis?

 Protein synthesis is the transcription and

translation of specific parts of DNA
(namely GENE) to form proteins.
 The following is a brief overview of how a
gene (a section of the DNA molecule)
serves as a template for the synthesis of a
protein. The process can be split into two
phases. Transcription occurs first, followed
by translation.
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 Steps in Translation
 mRNA brings the copied DNA code
from the nucleus to the cytoplasm
 mRNA attaches to one end of a
ribosome; called initiation
 tRNA’s attach the correct amino
acid floating in the cytoplasm to
themselves
 tRNA with its attached amino acid
have 2 binding sites where they join
the ribosome
 The tRNA anticodon “reads” &
temporarily attaches to the mRNA
codon in the ribosome
 Two amino acids at a time are linked
together by peptide bonds to make
polypeptide -chains (protein subunits);
called elongation
 Ribosomes) move along the mRNA
strand until they reach a stop codon
(UAA, UGA, or UAG); called
termination

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 Translation

 Translation is the part of protein synthesis where

the ribosomes in the cytoplasm use transfer
RNA (tRNA) to attach to the mRNA and translate
the bases into amino acids. tRNA molecules
bring the specified amino acids that the
ribosome links together to make a protein.
 http://vcell.ndsu.nodak.edu/animations/translatio
n/movie.htm

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 Amino Acids
 20 exist
 Linked together in a process called protein synthesis in the
cytoplasm to make polypeptides (subunits of proteins)
 DNA contains the instructions for making proteins but is too large
to leave the nucleus
 Three consecutive bases on DNA called a triplet (e.g. TCG, ATG,
ATT)
 mRNA codon table tells what 3 bases on mRNA code for each
amino acid (64 combinations of 3 bases)
 Methionine (AUG) on mRNA is called the start codon because it
triggers the linking of amino acids
 UAA, UGA, & UAG on mRNA signal ribosomes to stop linking
amino acids together

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 Latihan
(Tuliskan dalam Buku Catatanmu):
Diketahui suatu gen memiliki info sebagai
berikut :
5’– GACTTAGATACCGAGGTAATAAATG-3’

a.Buatlah pasangannya menjadi gen yang utuh

b.Buatlah mRNA darinya (hasil transkripsi)
c.Buatlah susunan asam amino dari protein yang terbentuk
(hasil translasi)
d.Jika basa T urutan ke-4 rusak akibat sinar radiasi,
tentukanlah rutan protein yang diberntuknya
e.Jika basa T urutan kelima rusak
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f.Jika basa A urutan ke-enam rusak
 Tugas Resensi Buku
 Carilah dalam literatur;
 bagaimana Ekspresi Genetik dikendalikan oleh tubuh
 Dituliskan dalam Buku Catatan

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 Ukuran berat molekul gen adalah bp (base
pair)
 gen dengan 100 bp, artinya ada 100
pasang basa nukleat

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