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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569 bp a salam m sofro .Human Genome Size • NUCLEAR GENOME * 23 pairs of chromosomes 2 x ( 3 x 109 b. 16.p)  2 meters DNA / Cell * 2 x ( 3 x 1012 cells) meters DNA in human body  8.000 x earth to moon • MITOCHONDRIAL GENOME * circular.

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

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

• The ribosome moves from codon to codon along the mRNA. a release factor binds to the stop codon. At the end. a salam m sofro . terminating translation and releasing the complete polypeptide from the ribosome. Amino acids are added one by one. 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 . UAG.Three nonsense codons (UAA.

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 = 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. a salam m sofro .

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

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

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 .

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

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

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purine to other purine) – Transversion (pyrimidine to purine or purine to pyrimidine) a salam m sofro .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.

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

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

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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. 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 .Model of gene expression in prokaryotes • One cistron.

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

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