Protein synthesis

(Translation of mRNA)

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Teaching aims
• By the end of the lecture:
– students are expected to understand
the molecular mechanism of translation
or protein synthesis
– students are expected to understand
the regulation of gene expression

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Core topics



Overview
Translation of mRNA
Mutation
Regulation of gene expression

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Transfer of genetic information
• Replication of DNA
 transmission of genetic information
from parental cell to its daughter cells
• Transcription of DNA
 transmission of genetic information
from DNA to RNA
• Translation of RNA (polypeptide/protein
biosynthesis)
 transmission of genetic information
from RNA to polypeptide/protein
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000 x earth to moon • MITOCHONDRIAL GENOME * circular.569 bp a salam m sofro .p)  2 meters DNA / Cell * 2 x ( 3 x 1012 cells) meters DNA in human body  8. 16.Human Genome Size • NUCLEAR GENOME * 23 pairs of chromosomes 2 x ( 3 x 109 b.

One of the most important stages in RNA processing is RNA splicing. may be interrupted by stretches of noncoding DNA. a salam m sofro . In many genes. or "exons". the DNA sequence coding for proteins. called "introns".

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The ribosome proceeds to the elongation phase of protein synthesis. During this stage. a salam m sofro .Translation of RNA • The ribosome binds to the mRNA at the start codon (AUG) that is recognized only by the initiator tRNA. complexes. sequentially bind to the appropriate codon in mRNA by forming complementary base pairs with the tRNA anticodon. composed of an amino acid linked to tRNA.

Amino acids are added one by one. a salam m sofro .• The ribosome moves from codon to codon along the mRNA. At the end. terminating translation and releasing the complete polypeptide from the ribosome. a release factor binds to the stop codon. translated into polypeptidic sequences dictated by DNA and represented by mRNA.

Codon • • • • • • Three-letter code words ( a triplet code) Unambiguous Non-overlapping Without punctuation Universal Can be found either in DNA (sense strand) and mRNA The collection of codons (64) makes up the genetic code a salam m sofro .

UGA) do not code for specific amino acid and are utilized as termination signal a salam m sofro .Three nonsense codons (UAA. UAG.

A = adenine G = guanine C = cytosine T = thymine U = uracil DNA transfers information to mRNA in the form of a code defined by a sequence of nucleotides bases. During protein synthesis. ribosomes move along the mRNA molecule and "read" its sequence three nucleotides at a time (codon) from the 5' end to the 3' end. a salam m sofro .

and then pairs with a sequence of three complementary nucleotides carried by a particular tRNA (anticodon). a salam m sofro .• Each amino acid is specified by the mRNA's codon. • Since RNA is constructed from four types of nucleotides. there are 64 possible triplet sequences or codons (4x4x4).

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The genetic code is said to be degenerate.• Three of these possible codons specify the termination of the polypeptide chain.   a salam m sofro . That leaves 61 codons to specify only 20 different amino acids. They are called "stop codons“ (nonsense codons). most of the amino acids are represented by more than one codon. Therefore.

Amino acids specified by each codon sequence on mRNA Ala: Alanine Cys: Cysteine Asp: Aspartic Glu: Glutamic acid acid Phe: Phenylalanine Gly: Glycine Ile: His: Histidine Isoleucine Lys: Lysine Met: Leu: Leucine Methionine Asn: Asparagine Pro: Proline Gln: Glutamine Arg: Arginine Ser: Serine Thr: Threonine Val: Valine Trp: Tryptophane a salam m sofro Tyr: Tyrosisne .

b and c for every amino acid incorporated into the protein being made): a. Elongation of the polypeptide chain (one repetition of the steps a.Protein translation takes place by the following steps 1. Formation of the initiation complex 2. translocation 3. peptide bond formation c. binding of aminoacyl-tRNA b. Termination a salam m sofro .

Remember ! • Proteins are polypeptides made up of individual amino acids linked together. and • Nucleic acids are polynucleotides made up of individual nucleotides linked together. a salam m sofro . • Carbohydrates are polysaccharides made up of individual monosaccharides linked together.

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Mutations • Result when changes occur in the nucleotide sequence  may not occur in the template strand but appear after replication • Some mutations occur by base substitution  single base changes (point mutations): – Transitions (pryrimidine to other pyrimidine. purine to other purine) – Transversion (pyrimidine to purine or purine to pyrimidine) a salam m sofro .

• Single base changes in DNA sequence followed by changes in mRNA molecules may have one of several effects when translated into protein: – No detectable effect  silent mutation – Missense effect  missense mutation – Appearance of nonsense codon that result in premature termination of polypeptide chain being synthesized  nonsense mutation a salam m sofro .

• Substitution of amino acids in protein causes missense mutations (illustration on Hemoglobin molecule): – Acceptable missense mutations • Hb Hikari: AAA or AAG (lys) to AAU or AAC (asp) • Hb E: GAA or GAG (glu) to AAA or AAG (lys) – Partially acceptable missense mutations • Hb S: GAA or GAG (glu) to GUA or GUG (val) – Unacceptable missense mutations • Hb M: Hb (Fe2+) to met Hb (Fe3+) a salam m sofro .

two. three or multiples nucleotides a salam m sofro .• Frameshift mutations result from deletion or insertion of nucleotides  generates altered mRNAs – May be one.

Regulation of gene expression a salam m sofro .

• In bacteria & viruses: – Alteration of gene expression is required by organism to adapt to environmental changes  involves interaction of specific binding proteins with various regions of DNA in the immediate vicinity of the transcription start site a salam m sofro .

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alteration of gene expression also involves tissue specific expression. metals & chemicals. gene amplification. regulation by hormones. posttranscriptional modification a salam m sofro . gene rearrangement.• In eukaryotes: – in addition to those proteins.

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Type of responses to a regulatory signal • Type A: increased rate of gene expression that is dependent upon the continued presence of inducing signal • Type B: increased rate of gene expression that transient even in the continued present of regulatory signal • Type C: increased rate of gene expression that persist indefinitely even after the termination of the signal a salam m sofro .

Type of gene regulation • Positive regulation: – The expression of genetic info is quantitatively increased by the presence of a specific regulatory element (the molecule is positive regulator) • Negative regulation: – the expression of genetic info is diminished by the presence of a specific regulatory element (the molecule is negative regulator) a salam m sofro .

one subunit concept instead of one gene one enzyme concept (cistron is the smallest unit of gene expression.Model of gene expression in prokaryotes • One cistron. coding for the structure of the subunit of a protein molecule) • Inducible genes: their expression increases in response to an inducer • Constitutive genes: their expression is reasonably constant (not known to be subject to regulation) a salam m sofro .

The earliest level of regulation is at DNA level during transcription a salam m sofro .

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