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    Translation

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    Anne Gabrielle Marapon Agsunod
    Translation is the process where mRNA is used as a template to synthesize polypeptide chains through the joining of amino acids. It involves three main steps - initiation, elongation, and termination. In initiation, initiation factors help assemble the ribosomal subunits and initiator tRNA on the start codon. In elongation, elongation factors and tRNAs sequentially add amino acids specified by mRNA codons to form the polypeptide chain. Termination occurs when release factors recognize stop codons and release the complete polypeptide chain.

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    Translation

    Uploaded by

    Anne Gabrielle Marapon Agsunod
    47 views44 pages
    Translation is the process where mRNA is used as a template to synthesize polypeptide chains through the joining of amino acids. It involves three main steps - initiation, elongation, and termination. In initiation, initiation factors help assemble the ribosomal subunits and initiator tRNA on the start codon. In elongation, elongation factors and tRNAs sequentially add amino acids specified by mRNA codons to form the polypeptide chain. Termination occurs when release factors recognize stop codons and release the complete polypeptide chain.

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    Translation is the process where mRNA is used as a template to synthesize polypeptide chains through the joining of amino acids. It involves three main steps - initiation, elongation, and termination. In initiation, initiation factors help assemble the ribosomal subunits and initiator tRNA on the start codon. In elongation, elongation factors and tRNAs sequentially add amino acids specified by mRNA codons to form the polypeptide chain. Termination occurs when release factors recognize stop codons and release the complete polypeptide chain.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    47 views44 pages

    Translation

    Uploaded by

    Anne Gabrielle Marapon Agsunod
    Translation is the process where mRNA is used as a template to synthesize polypeptide chains through the joining of amino acids. It involves three main steps - initiation, elongation, and termination. In initiation, initiation factors help assemble the ribosomal subunits and initiator tRNA on the start codon. In elongation, elongation factors and tRNAs sequentially add amino acids specified by mRNA codons to form the polypeptide chain. Termination occurs when release factors recognize stop codons and release the complete polypeptide chain.

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    TRANSLATION

    Translation
    • Process wherein polypeptide chains are synthesized
    • Uses mRNA as template which determines the order
    in which different amino acid residues are to be
    joined
    Translation: Prokaryotes vs.
    Eukaryotes
    THE
    TRANSLATION
    mRN
    A• Template for translation
    • Codon
    • Three adjacent ribonucleotides along the mRNA
    which correspond to a specific amino acid
    The Genetic Code
    • The correspondence between codons and amino
    acids
    The Genetic Code: Properties
    • Universal (almost)

    Codon Universal code Unusual code Occurence

    UGA stop Trp Mycoplasma, mitochondria

    UGA stop Cys Euplotes

    UAA, UAG stop Gln Various protozoa

    CUG Leu Thr Yeast mitochondria

    CUG Leu Ser Candida


    The Genetic Code: Properties
    • Degenerate
    • Some amino acids are coded by more than one
    codon
    • e.g. Phe is coded by UUU and UUC
    • Contiguous
    • Codons do not overlap and have no spacers in
    between
    • Unambiguous
    • Each codon specifies only one amino acid
    Prokaryotic
    mRNA
    • Contains the Shine-Dalgarno Sequence
    • Found near the 5’-end of the mRNA
    • Sequence responsible for proper positioning of the
    mRNA in the ribosome
    • Complementary to a portion of the 16S rRNA in
    the 30S (small) ribosomal unit
    • Consensus sequence: 5’-AGGAGGU-3’
    tRNA
    • Carries the amino acid
    to the translation
    machinery
    • Contains the
    anticodon that is
    complementary to the
    codon in the mRNA
    • Has a cloverleaf
    structure
    Parts of tRNA
    • Acceptor stem
    • the 5’-CCA-3’ terminal
    that binds the amino
    acid through an
    ester bond
    • Anticodon arm
    • contains the
    anticodon that is
    complementary to the
    codon it recognizes
    in the mRNA
    Parts of tRNA
    • D or DHU arm
    • TΨCG arm
    • contribute
    important
    interactions for
    the overall folding
    of tRNA molecules
    • Rich in unusual
    nitrogenous bases
    Tertiary Structure of tRNA
    • Inverted L-shaped structure
    • Formed by 9 H-bonds mainly between the unusual
    bases in the variable regions
    The Genetic Code

    • 64 codons – 3 stop codons = 61 tRNAs needed


    The Wobble Hypothesis
    • Rules of base pairing
    are relaxed in the third
    position of the codon
    • Reduces the number
    of tRNAs needed by
    the cell
    • Enables one tRNA to
    read two or three
    codons
    The Wobble Hypothesis
    Ribosomes
    • Composed of:
    • Large subunit
    • contains the peptidyl transferase center -
    catalyzes peptide bond formation
    • factor-binding center
    • Small subunit
    • contains the decoding center: narrow channels
    where the mRNA enters and exits
    Ribosomes
    • A site
    • Aminoacyl tRNA
    binding site
    • P site
    • Peptidyl tRNA
    binding site
    • E site
    • Exit site
    Sedimentation Coefficient
    • a measure of the rate of sedimentation in an
    ultracentrifuge of a molecule suspended in a less
    dense solvent.
    • measured in Svedberg units, S.
    • S values are not additive.
    Eukaryotic Ribosomes
    • Large subunit – 60S
    • 50 proteins + (28S + 5.8S + 5S) rRNA
    • Small subunit – 40S
    • 33 proteins + 18S rRNA
    Prokaryotic Ribosomes
    • Large subunit – 50S
    • 31 proteins + (23S + 5S) rRNA
    • Small subunit – 30S
    • 21 proteins + 16S rRNA
    PROKARYOTIC
    Amino Acid Activation

    Alberts et al 2008
    Amino Acid Activation
    Amino acid + ATP →
    amino acyl-AMP + PPiAmino acyl-AMP +
    tRNA → amino acyl-tRNA + AMP
    • Catalyzed by: amino acyl-tRNA
    synthetase
    • Eg. for the activation of alanine
    • Enzyme: alanine-tRNA
    synthetase
    • Activated form: alanine-tRNAAla
    Initiation Complex Formation
    • Initiator tRNAifmet
    • recognizes AUG (and GUG or
    UUG by wobble)
    • aminoacylated with met
    • met is formylated in its
    amino group to become
    N-formylmethionine
    (fmet)
    • f-met as temporary N-
    terminus of the peptide
    Initiation Complex Formation
    1. IF1 promotes dissociation of ribosomal subunits
    2. IF3 binds to 30S subunit to prevent its re-association
    with the 50S subunit
    Initiation Complex Formation
    3. mRNA binds to the 30S subunit
    • Shine-Dalgarno sequence as ribosome binding site
    Initiation Complex Formation
    4. IF2 (along with
    GTP) brings the
    initiator tRNA fmet
    to the 30S
    subunit.
    Initiation Complex Formation
    5. IF1 bound to the
    initiation complex
    • induces
    conformational
    change
    • allows the 50S
    subunit to attach
    using energy from
    GTP hydrolysis.
    • IF’s are then
    released.
    Elongation of Polypeptide
    1. Binding of the next charged amino acid
    • A-site as entry point
    • Facilitated by EF-Tu and requires hydrolysis of GTP
    • EF-Ts recharges EF-Tu with GTP
    2. Peptide bond formation
    • Polypeptide from the tRNA in the P-site forms a
    peptide bond with the amino acid attached to the
    tRNA in the A-site
    • Catalyzed by peptidyl transferase
    Elongation of Polypeptide

    EF-Tu

    Peptide bond formation


    catalyzed by
    peptidyl transferase
    Elongation of Polypeptide
    3. Translocation
    • Ribosome slides through the mRNA to read the
    next codon
    • Promoted by another elongation factor EF-G and
    requires GTP hydrolysis
    • Places empty tRNA in the E-site
    • tRNA with growing polypeptide chain now on the
    P-site
    Elongation of Polypeptide

    EF-G
    Termination of Translation
    1. Recognition of stop codons in the A-site
    • UAG, UGA, UAA
    2. Binding of release factor (RF) in the A-site
    • RF1 recognizes UAG and UAA;
    • RF2 recognizes UGA and UAA;
    • RF3 assists RF1 or RF2
    • Hydrolyzes the polypeptide chain from the tRNA
    3. Dissociation of the translation machinery
    Termination of Translation
    Transcription and Translation
    EUKARYOTIC
    Initiation Factors
    Prokaryotic Eukaryotic Function
    IF1, IF3 eIF3, eIF4C, eIF6 • Binding to ribosome
    subunits
    IF2 eIF4B, • Binding to mRNA
    eIF4F eIF2, • Initiator tRNA delivery
    eIF2B eIF5 • Displacement of other
    factors
    Translation
    Initiation
    • Prokaryotes
    • translation initiation complex is built directly on
    start codon
    • facilitated by the Shine-Dalgarno sequence
    • Eukaryotes
    • ribosomes indirectly locate start codon
    • there is no ribosome-binding site in eukaryotic
    mRNA
    • involves a scanning mechanism starting at the
    5’
    cap
    Translation Initiation
    • Scanning
    Mechanism
    Translation Initiation
    • Scanning Mechanism
    Translation
    Elongation
    • Elongation Factors:
    • eEF-1α (= EF-Tu)
    • eEF-1βγ (= EF-Ts)
    • eEF-2 (= EF-G)

    • no E-site in ribosome
    Protein Targeting in
    Eukaryotes
    • to target proteins into their respective location
    within or outside of the cell before they become
    biologically active
    • Involves:
    • signal peptide
    • in the N-terminal of the nascent protein
    • signal recognition particle (SRP)
    • recognizes signal peptide
    Events in Protein Targeting

    • Glycosylation pattern (step 6) control the final


    location of the protein.

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