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    The Genetic Code and Transcription: Flow of Genetic Information

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    Renz L. Salumbre
    The document discusses the flow of genetic information from DNA to protein. It covers the genetic code, including the discovery that it uses triplets of nucleotides, is degenerate, and universal across organisms. Key events included the discoveries by Nirenberg and others that codons specify amino acids. The document also discusses transcription, including the key parts of prokaryotic and eukaryotic transcription as well as post-transcriptional processing of eukaryotic pre-mRNA through capping, polyadenylation, splicing, and other mechanisms.

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    The Genetic Code and Transcription: Flow of Genetic Information

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    Renz L. Salumbre
    638 views10 pages
    The document discusses the flow of genetic information from DNA to protein. It covers the genetic code, including the discovery that it uses triplets of nucleotides, is degenerate, and universal across organisms. Key events included the discoveries by Nirenberg and others that codons specify amino acids. The document also discusses transcription, including the key parts of prokaryotic and eukaryotic transcription as well as post-transcriptional processing of eukaryotic pre-mRNA through capping, polyadenylation, splicing, and other mechanisms.

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    Genetics Lecture Handouts

    Original Title

    Gene Lecture 11 Transcription

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    The document discusses the flow of genetic information from DNA to protein. It covers the genetic code, including the discovery that it uses triplets of nucleotides, is degenerate, and universal across organisms. Key events included the discoveries by Nirenberg and others that codons specify amino acids. The document also discusses transcription, including the key parts of prokaryotic and eukaryotic transcription as well as post-transcriptional processing of eukaryotic pre-mRNA through capping, polyadenylation, splicing, and other mechanisms.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    638 views10 pages

    The Genetic Code and Transcription: Flow of Genetic Information

    Uploaded by

    Renz L. Salumbre
    The document discusses the flow of genetic information from DNA to protein. It covers the genetic code, including the discovery that it uses triplets of nucleotides, is degenerate, and universal across organisms. Key events included the discoveries by Nirenberg and others that codons specify amino acids. The document also discusses transcription, including the key parts of prokaryotic and eukaryotic transcription as well as post-transcriptional processing of eukaryotic pre-mRNA through capping, polyadenylation, splicing, and other mechanisms.

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    of 10

    Fundamental Genetics

    Lecture 9

    The Genetic Code


    and Transcription

    John Donnie A. Ramos, Ph.D.


    Dept. of Biological Sciences
    College of Science
    University of Santo Tomas

    Flow of Genetic Information

    1
    The Genetic Code
    ‰ Linear form (mRNA derived from DNA)
    ‰ Triplet codons (triplets of ribonucleotides coding for 1
    amino acid)
    ‰ Unambiguous (1 codon = 1 amino acid only)
    ‰ Degenerate ( 1 amino acid can be specified by several
    codons)
    ‰ Contains specific start and stop codons
    ‰ Commaless (no breaks once translation starts until the
    stop codon is reached)
    ‰ Non-overlapping (single reading frame)
    ‰ Universal (same ribonucleotide used by all organisms)

    The Discovery of the Genetic Code


    ‰ Francois Jacob and Jacques Monod
    (1961) – messenger RNA (mRNA)
    ‰ Sydney Brenner (1960s) – codon in
    triplets (minimal use of the 4 mRNA
    bases to specificy 20 aa) (43=64)
    ‰ Francis Crick – frameshift mutations
    alters the codons
    ‰ Mariane Manago and Severo Ochoa -
    polynucleotide phosphorylase
    (synthesis of RNA without template)-
    paved the way to the production of
    RNA polymeres in cell free-systems

    2
    The Discovery of the Genetic Code
    ‰ Marshall Nirenberg and J. Heinrich Matthaei (1661) – codons
    ‰ used cell-free protein synthesizing system and polynucleotide
    phosphorylase
    ‰ RNA Homopolymers (UUUUUU…, AAAAAAA…, CCCCCC…, GGGGG…)
    ‰ UUU (Phenylalanine)
    ‰ AAA (Lysine)
    ‰ CCC (Proline)
    ‰ RNA Mixed Copolymers

    1A:5C (1/6 A: 5/6C)

    The Triplet Binding Assay


    ‰ Developed by M. Nirenberg and P. Leder (1964)
    ‰ Mimics the in vivo translation of proteins where a mRNA-tRNA-
    ribosome complex is formed when all three macromolecules are
    allowed to interact.

    3
    Repeating Copolymers
    ‰ Developed by Gobind Khorana (1960s)
    ‰ Synthetic long RNAs with repeating sequences

    Results of Repeating Copolymers

    4
    The Universal Genetic Code

    ‰ Degeneracy
    ‰ Wobble
    Hypothesis
    ‰ Start codon
    (N-formylmethionine)

    ‰ Termination
    codons
    ‰ Universal
    Viruses
    Bacteria
    Archaea
    Eukaryotes

    Exceptions to the Universal Code

    5
    Transcription
    ‰ Uses DNA as a template
    ‰ Catalyzed by RNA polymerase
    (holoenzyme of 500 kD)
    ‰ αββ’σ subunits
    ‰ Sense strand / template strand –
    DNA strand used as a template
    for transcription
    ‰ Promoter region – DNA sequence
    recognized by σ factor to initiate
    transcription (60 bases).
    (upstream of a gene)
    ‰ TATA box (Pribnow box) –
    TATAAT sequence
    ‰ Sigma factor (σ70, σ28, σ32, σ54)

    Transcription
    ‰ RNA polymerase don’t need
    primers
    ‰ Elongation in 5’ to 3’ direction
    ‰ Rate in E coli: 50 bases/sec at
    37°C
    ‰ Termination is a function of rho
    (ρ) factor – hexameric protein
    interacting with the end of a
    gene
    ‰ Polycistronic mRNA – bacterial
    mRNA containing information for
    the synthesis of proteins of
    related function
    ‰ Monocystronic mRNA –
    eukaryotic mRNA containing
    information for a single protein.

    6
    Eukaryotic Transcription
    ‰ Features of eukaryotic transcription different from prokaryotic
    transcription:
    ‰ Transcription inside the nucleus under the direction of 3 different
    RNA polymerases

    ‰ Presence of protein factors (promoters, enhancers, etc.) binding to


    the upstream portion of a gene (cis-acting elements) during
    initiation step.
    ‰ Presence of post-transcriptional regulation.

    Cis -acting Elements


    ‰ TATA Box (Goldberg-Hogness Box)
    ‰ Located 30 bases upstream from the start of
    transcription (-30)
    ‰ Consensus sequence: TATAAAA
    ‰ Facilitates denaturation of helix because it is AT-
    rich region

    ‰ CAAT Box
    ‰ Located 80 bases upstream from the start of
    transcription (-80)
    ‰ Consensus sequence: GGCCAATCT
    ‰ Influence the efficiency of the promoter

    7
    Trans -acting Factors
    ‰ Transcription factors – facilitates template
    binding during the initiation of transcription
    ‰ Example:
    ‰ TFIID (TATA-binding protein or TBP) – binds to
    TATA-box

    Post-transcriptional Processing
    ‰ 7-methylguanosine cap (7mG)
    ‰ Protection from nucleases
    ‰ Role mRNA transport across the
    nuclear membrane
    ‰ 3’ cleavage site:
    ‰ AAUAAA
    ‰ Failure of 3’ cleavage results to
    absence of poly A tail
    ‰ Split genes – contains intervening
    sequences

    8
    RNA Splicing

    ‰ Ribozyme – RNA with


    catalytic activity
    ‰ Self-excision process –
    process of RNA splicing or
    intron removal.
    ‰ Transesterification –
    interaction between
    guanosine and the
    transcript.
    ‰ 2 successive
    transesterification processes

    The Spliceosome

    ‰ Alternative splicing
    ‰ Small nuclear ribonucleoproteins
    (snRNP or snurps) – bonds to GU
    or AG sites of introns
    ‰ 2 transesterification processes
    ‰ Snurps form a loop (lariat) in the
    branch point region
    ‰ Produces isoforms of proteins

    9
    RNA Editing

    ‰ Substitution editing
    ‰ changes in the nucleotide bases of a given mRNA
    ‰ Common in mitochondrial RNA and chloroplast RNA
    ‰ Example: Apoliprotein B (Apo B) – C to U change CAA
    to UAA
    ‰ Insertion / deletion editing
    ‰ addition or removal of nucleotide sequences
    ‰ Common in mitochondrial RNA or guide RNA (gRNA)

    10

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