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Ch. 17: From Gene to Protein DNA RNA Protein Protein Synthesis: overview One gene-one enzyme hypothesis (Beadle and Tatum) One gene-one polypeptide (protein) hypothesis Transcription: synthesis of RNA under the direction of DNA (mRNA) Translation: actual synthesis of a polypeptide under the direction of mRNA The Triplet Code The genetic instructions for a polypeptide chain are ‘written’ in the DNA as a series of 3-nucleotide ‘words’ Reading Frame w/ Dictionary Codons code for Amino Acid ‘U’ (uracil) replaces ‘T’ in RNA Comparison of DNA and RNA Types of sugar nitrogenous bases single vs. double strand function DNA Deoxy Ribose G,C,T,A DS Replication RNA Ribose G,C,A,U SS Proteins Processes from Gene to Protein Transcription Synthesis of mRNA Translation Synthesis of Protein Transcription, I Promoter region on DNA: where RNA polymerase attaches and where initiation of RNA begins RNA polymerase: pries DNA apart and hooks RNA nucleotides together from the DNA code Terminator region: sequence that signals the end of transcription Transcription unit: stretch of DNA transcribed into an RNA molecule Transcription II Initiation~ transcription factors mediate the binding of RNA polymerase to an initiation sequence (TATA box) Elongation~ RNA polymerase continues unwinding DNA and adding nucleotides to the 3’ end Termination~ RNA polymerase reaches terminator sequence Eukaryote Cells vs. Prokaryote Initiation of transcription RNA polymerase Prokaryotes bind to RNA Poly directly. Eukaryotes bind to transcription factors that bind to DNA. promoters (TATA Box) transcription factors Transcription initiation complex mRNA modification 1) 5’ cap: modified guanine; protection; recognition site for ribosomes 2) 3’ tail: poly(A) tail (adenine); protection; recognition; transport 3) RNA splicing: exons (expressed sequences) kept introns (intervening sequences) are spliced out forming a spliceosome What are they????? Introns Exons RNA Splicing •Introns: intervening sequence. Non coding •Exon: translates to Amino Acids sequence •Spliceosome/Ribozymes Small nuclear ribonucleoproteins Evolutionary Importance?? Alternative RNA Splicing Gene gives rise to different proteins depending on which segments are exons during RNA processing Potentially new proteins w/ new functions Increase chance of crossing over b/w genes, increase genetic recombination Translation, I RNA to Protein mRNA from nucleus is ‘read’ along its codons by tRNA’s anticodons at the ribosome tRNA anticodon (nucleotide triplet); amino acid Translation, II rRNA site of mRNA codon & tRNA anticodon coupling P site holds the tRNA carrying the growing polypeptide chain A site holds the tRNA carrying the next amino acid to be added to the chain E site discharged tRNA’s Translation, III Initiation~ union of mRNA, tRNA, small ribosomal subunit; followed by large subunit Elongation~ •codon recognition •peptide bond formation •translocation Termination~ ‘stop’ codon reaches ‘A’ site Polyribosomes: translation of mRNA by many ribosomes (many copies of a polypeptide very quickly) Role of RNA Coupling in Bacterial DNA Transcription Factors Compartmentalization Bacteria couple Transcription and translation Signals for Specific Destinations Free Ribosomes Proteins used in cytosol Bound Ribosomes Attached to ER Endomembrane System Secreted from Cell (Insulin) Polypeptide marked by signal peptide (20a.a at leading end, 5’ cap modified) Recognized by Signal Recognition Particle (SRP) Mutations: genetic material changes in a cell Point mutations…. Changes in 1 or a few base pairs in a single gene Base-pair substitutions: silent mutations: no effect on protein missense : ∆ in sequence, a different amino acid (different protein) Nonsense: ∆ to a stop codon causes a nonfunctional protein Base-pair insertions or deletions: additions or losses of nucleotide pairs in a gene; alters the ‘reading frame’ of triplets~frameshift mutation Mutagens: physical and chemical agents that change DNA “What would happen if…..” A mutation changed a start codon to some other codon? A mutation changed a stop codon to some other codon? A mutation changed one amino acid within a polypeptide chain? Exploring a Gene