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The Genetic

&
Code

Translation Process
(Protein Biosynthesis)

Department of Biochemistry and Molecular Biology
School of Medicine, Tarumanagara University

October 2010

The Genetic Code
• The genetic code is the set of rules by which information encoded in genetic
material (DNA or RNA sequences) is translated into proteins (amino acid
sequences) by living cells.
• mRNA molecules have no affinity for amino acids and, therefore, that the translation
of the information in the mRNA nucleotide sequence into the amino acid sequence of
a protein requires an intermediate adapter molecule.
• This adapter molecule (tRNA) must recognize a specific nucleotide sequence on the
one hand as well as a specific amino acid on the other.
• Twenty amino acids are required for the synthesis of the cellular proteins; thus, there
must be at least 20 distinct codons that make up the genetic code.
• Since there are only 4 different nucleotides in mRNA, each codon must consist of
more than a single purine or pyrimidine nucleotide. Codons consisting of two
nucleotides each could provide for only 16 (42) specific codons, whereas codons
of three nucleotides could provide 64 (43) specific codons.

• The mRNA transcript is a linear sequence of
nucleotides carrying genetic information and
it is single-stranded.
• Every three bases of mRNA (a triplet) specifies
an amino acid to be added to a growing
polypeptide chain; the relationship between
the triplets and the corresponding amino acids
is the genetic code.
• Each base triplet of mRNA is called a codon.
The genetic code is nearly universal for all
forms of life.

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Features of the Genetic Code .

• The order of the codons in the mRNA determines the linear sequence of amino acids in the protein.MARSHALL Nirenberg (1961) Relationship between mRNA and the Protein Product • The order of the codons in the mRNA determines the sequence in which amino acids are added to the growing polypeptida chain.>Produce stop kodon InsertionDeletion- > Addition one or more bases > Loss one or more bases Example CGA . TYPE OF MUTATIONS TYPE Point SilentMissense- Description > A Single base change > A change that specifies the same amino > Different Amino Nonsense .-> CGG Arg->Arg CGA->CCA Arg -> Pro CGA->UGA Arg > Stop .

all the components needed for translation come together. . • In the first step of the process.Translation of mRNA (Protein Synthesis) • Translation is the process by which the nucleotide sequence of mRNA is converveted to the amino acid sequence of a polypeptide.

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mRNA .

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(Kozak Scanning) .

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Formation of aminoacyl-tRNA Amino acid activation aminoacyl-tRNA synthetase .

which couples a particular amino acid to its corresponding tRNA. whose anticodon forms base pairs with the appropriate nucleotide .The genetic code is translated by means of two sequential "adaptors". The first adaptor is the aminoacyltRNA synthetase enzyme. the second adaptor is the tRNA molecule.

A polypeptide chain grows by the stepwise addition of amino acids to its carboxylterminal end. The formation of each peptide bond is energetically favorable because the .The incorporation of an amino acid into a protein.

Decoding an mRNA molecule Each amino acid added to the growing end of a polypeptide chain is selected by complementary base-pairing between the anticodon on its .

A comparison of the structures of procaryotic and eucaryotic .

The elongation phase of protein synthesis on a .

Kinetic proofreading selects for the correct tRNA molecule on the ribosome .

The final phase of protein synthesis: The binding of release factor to stop codon terminate .

The initiation phase of protein synthesis in eucaryotes .

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Recognition of the codon by the anticodon .

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Translation initiation pathway in E. coli ribosom .

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4G. . 4E.Translational initiation pathway in eukaryotes. Several of the initiation factors: (4A. and 4H) have been left out of the 48S initiation complex for clarity . 4B. Initiation factors are represented by colored rectangles where they are first implicated in the pathway and by circles of the same color thereafter.

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• The flow of genetic information follows the sequence DNA → RNA → protein.
• The genetic information in the structural region of a gene is transcribed into an RNA
molecule such that the sequence of the latter is complementary to that in the DNA.
• Ribosomal RNA (rRNA), transfer RNA (tRNA), and messenger RNA (mRNA), are
directly involved in protein synthesis; miRNAs regulate mRNA function at the level
of translation and/or stability.
• The information in mRNA is in a tandem array of codons, each of which is 3 nt long.
• mRNA is read continuously from a start codon (AUG) to a termination codon
• The open reading frame, or ORF, of the mRNA is the series of codons, each specifying
a certain amino acid, that determines the precise aa sequence of the protein.
• Protein synthesis, like DNA and RNA synthesis, follows the 5' to 3' polarity of mRNA
and can be divided into three processes: initiation, elongation, and termination.

• Mutant proteins arise when single-base substitutions result in codons that specify a
different amino acid at a given position, when a stop codon results in a truncated protein,
or when base additions or deletions alter the reading frame, so different codons are read.
• A variety of compounds, including several antibiotics, inhibit protein synthesis by
affecting one or more of the steps involved in protein synthesis.

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• Anisomycin blocks the peptidyl transferase reaction on ribosomes • α-Amanitin blocks mRNA synthesis by binding preferentially to RNA polymerase II . • Chloramphenicol blocks the peptidyl transferase reaction on ribosomes • Erythromycin blocks the translocation reaction on ribosomes • Rifamycin blocks initiation of RNA chains by binding to RNA polymerase (prevents Acting Only on Procaryotes and Eucaryotes RNA synthesis) • Puromycin causes the premature release of nascent polypeptide chains by its addition to growing chain end. • Actinomycin D binds to DNA and blocks the movement of RNA polymerase Acting OnlyRNA on synthesis) Eucaryotes (prevents • Cycloheximide blocks the translocation reaction on ribosomes.Inhibitors of Protein or RNA Synthesis Acting Only on Procaryotes* • Tetracycline blocks binding of aminoacyl-tRNA to A-site of ribosome • Streptomycin prevents the transition from initiation complex to chainelongating ribosome and also causes miscoding.

. sedangkan di pihak lain sel dapat tumbuh dan berdiferensiasi secara normal. regulasi ekspresi gen terjadi dalam berbagai tingkat.REGULASI EKSPRESI GEN • Ekspresi gen merupakan keseluruhan proses. • Untuk mencapai tujuan tersebut . di mana informasi genetik yang disimpan dalam DNA diubah menjadi produk fungsional gen yaitu RNA atau protein. • Tujuan regulasi ekspresi gen adalah agar ekspresinya terjamin secara akurat baik spatial maupun temporal. sehingga di satu pihak sel dapat beradaptasi terhadap perubahan lingkungan.

or . (4) selecting which mRNAs in the cytoplasm are translated by ribosomes (translational control). (3) selecting which completed mRNAs in the cell nucleus are exported to the cytoplasm (RNA transport control).REGULASI EXPRESSI GENE (1) controlling when and how often a given geneis transcribed (transcriptional control). inactivating. (5) selectively destabilizing certain mRNA molecules in the cytoplasm (mRNA degradation control) (6) selectively activating. (2) controlling how the primary RNA transcript is spliced or otherwise processed (RNA processing control).

Base Pairing between mRNA and the 16S rRNA Helps Select the Translational Initiation Site c. (1961) b. Polypeptide Synthesis Proceeds from N-Terminus to C-Terminus (Howard Dintzis. Ribosomes Read mRNA in the 5’ → 3’ direction C. Chain Elongation Occurs by the Linkage of the Growing Polypeptide to the Incoming tRNA’s Amino Acid Residue Chain Initiation a.Polypeptide Synthesis: An Overview a. Eukaryotic Initiation Is Far More Complicated than That of Prokaryotes . Prokaryotic Initiation Is a Three-Stage Process that Requires the Participation of Soluble Protein Initiation Factors d. Active Translation Occurs on Polyribosomes d. fMet Is the N-Terminal Residue of Prokaryotic Polypeptides b.

Translational initiation pathway in eukaryotes. Initiation factors are represented by colored rectangles where they are first implicated in the pathway and by circles of the same color thereafter. 4E. 4B. The higher order complexes are hypothetical. Several of the initiation factors: (4A. 4G. and 4H) have been left out of the 48S initiation complex for clarity .

Signal Peptides Are Removed from Nascent Proteins by a Signal Peptidase C.Collagen Assembly Requires Chemical Modification . Polyproteins D. Proteolytic Cleavage B.POSTTRANSLATIONAL MODIFICATION A. Covalent Modification: .

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ex:.Additional PTMs • Nitrosylation • Glycation • ADP-ribosylation • O-GlcNAc • Lipidation.Prenyl groups. Farnesylation .Glycophosphatidyl inositol (GPI) membrane protein .

Protein Folding • Protein disulfide isomerase (PDI) • Molecular Chaperones: • heat shock proteins (Hsp). coli). coli) • Hsp10 proteins (GroES in E. The X-ray structure of the GroEL–GroES–(ADP)7 chaperonin complex . ex: HSP70 • Chaperonins: • Hsp60 proteins (GroEL in E.

in an ATP-dependent manner. It often works in collaboration with other chaperones. may help proteins fold to the native state OR Hsp70 may ‘transfer’ non-native proteins to other chaperones for folding (e. and has numerous other functions in the cell apart from assisting de novo protein folding..g. especially Hsp40 . chaperonins)  Hsp70 is also important during cellular stresses (thermotolerance).Hsp70 in de novo protein biogenesis  Hsp70 is believed to bind and stabilize nascent polypeptides early in their synthesis--preventing misfolding and aggregation  Hsp70 binding and release.