From Gene to Protein Chapter 17 Gene Expression The process by which DNA directs the synthesis of proteins 2 stages: transcription and translation Detailed flow of info from gene to protein Explain how genetic mutations affect organisms through their proteins Central Dogma (Francis Crick) DNA transcribed RNA translated Proteins link DNA to phenotype protein Decoding Genes to Phenotype Archibald Garrod Inborn error of metabolism Inherited disease from inability to make a particular enzyme Suggested that genes dictate phenotype through enzymes that catalyze specific reactions George Beadle and Edward Tatum One gene – one enzyme hypothesis A single gene specifies synthesis of a single enzyme in the body Shared Nobel Prize Additional Scientists Not all proteins are enzymes and many proteins are 1+ polypeptides Revised to one gene – one polypeptide Each gene codes for 1 polypeptide of a protein From DNA to Protein Genes instruct, but don’t build Nucleotides and amino acids are different ‘languages’ RNA connects them Transcription: same language Translation: different language Occurs in all organisms DNA Sugar is deoxyribose Has –H RNA Sugar is ribose Has -OH Bases are A,C, G, and T Bases are A, C, G, and U Double-stranded helix Single-stranded Only in nucleus Not confined to nucleus Modified only by mutations Lots of processing and 1 type modifications 3 types (mRNA, tRNA, rRNA) Reviewing DNA and RNA Summary of Protein Synthesis Genes determine the sequence of bases along an mRNA Only template strand is transcribed Similar to DNA replication mRNA is complementary to template strand Non-template strand is ‘identical’ to mRNA = coding strand Sequence of mRNA as codons translated to amino acids Transcription In the nucleus Initiation RNA polymerase binds to promoter Many work at once = efficiency Elongation Builds 5’ 3’ = downstream Unstable complex so mRNA immediately released and DNA rejoins Termination Terminator reached = releasing transcript (pre-mRNA) mRNA Processing Before mRNA leaves the nucleus Alteration of 5’ and 3’ ends 5’ cap, modified G, directs ribosome attachment for protein synthesis Poly-A tail, addition of 5-250 adenines (A), inhibits degradation as leaves nucleus RNA splicing Pre-mRNA transcript contains exons and introns RNA sequences and DNA sequences that encode them Spliceosomes splice out introns and rejoin exons = true mRNA Translation In the cytoplasm within ribosomes mRNA as codon message from DNA Translated by tRNA Anticodons and amino acid ends Ribosomes facilitate addition of tRNA to mRNA 3 steps like transcription Ribosomes Facilitate coupling of tRNA anticodons and mRNA codons Large and a small subunits Functional when subunits join with mRNA 3 unique binding sites facilitate More than 1 can bind to a single mRNA Initiation Small ribosome subunit binds to mRNA and moves to start codon 1st tRNA enters the P site carrying the AA met Anticodon is what? Large subunit binds Initiation factors facilitate and GTP supplies energy Elongation 2nd tRNA molecule enters the A site Once matched the growing polypeptide chain binds to the new AA Ribosome shifts 5’ 3’, changing P site tRNA to E site, A site to P site, and freeing A site = translocation Termination Stop codon sequence that signifies the end of a polypeptide chain Sequences are UAG, UAA, and UGA Don’t code for AA’s Polypeptide cleaved from last tRNA (P site) and leaves the ribosome Folds into quaternary structure = protein Decoding Codons Only 4 nucleotide bases to specify 20 amino acids Genetic instructions are based on codons 42 = 16 (not enough); 43 = 64 (plenty) Demonstrates redundancy, but not ambiguity 3rd base is wobble base Nearly universal across species Mutations Changes to the genetic information of a cell, or virus Ultimate source of diversity because ultimate source of new genes Chapter 15 was large scale mutations which effect long DNA segments What were the 4 types? Point mutations change single nucleotides and causes changes in single specific codons Base-pair substitution Frameshift mutations result from altering the reading frame Base-pair insertions/deletions Base-pair Substitutions Replaces 1 nucleotide pair with another Effect depends on particular codon change and location Silent mutations occur at wobble bases so have no effect on the encoded protein Missense mutations change 1 AA to another Nonsense mutation codes for a stop codon rather than an AA Stops translation early Can result in wrong AA being added, abnormal protein shape, or shortened proteins Alters active site and can make non-functional Frameshift Mutations Insertions and deletions add or remove nucleotide pairs of a gene Often more deleterious when changes don’t occur in groups of three Any downstream bases will be affected Almost always causes protein to be nonfunctional THE CAT ATE THE RAT THE ATA TET HER AT (delete C) THE CAT GAT ETH ERA T (insert G)