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Halloween pets? Student Assessment of Learning Gains (SALG) website Last Night’s Homework 1. Compare and Contrast DNA Polymerase (like Taq) versus RNA Polymerase. What's the same, what's different? 2. Taq polymerase starts it's work at a primer when you do PCR, how does RNA polymerase figure out where to start it's work during transcription? 3. The coding region of a gene is essentially the exons. The non-coding region of a particular gene is everything else. What gene are you studying in your research project? How many exons does it contain? If a mutation occurs in an intron can that cause your disease? What if a mutation occurred in the TATA box? Last Night’s Homework 1. Compare and Contrast DNA Polymerase (like Taq) versus RNA Polymerase. What's the same, what's different? 2. Taq polymerase starts it's work at a primer when you do PCR, how does RNA polymerase figure out where to start it's work during transcription? 3. The coding region of a gene is essentially the exons. The non-coding region of a particular gene is everything else. What gene are you studying in your research project? How many exons does it contain? If a mutation occurs in an intron can that cause your disease? What if a mutation occurred in the TATA box? In-class work The coding region of a gene is essentially the exons. The non-coding region of a particular gene is everything else. If a mutation occurs in an intron can that cause a disease? What if a mutation occurred in the TATA box? DNA molecule Gene 2 Gene 1 Gene 3 Vocabulary? DNA template strand TRANSCRIPTION mRNA Codon TRANSLATION Protein Amino acid Signal Gene Expression NUCLEUS Chromatin Chromatin modification DNA Gene available for transcription Gene Transcription RNA Exon Primary transcript Intron RNA processing Tail Cap mRNA in nucleus Transport to cytoplasm CYTOPLASM CYTOPLASM mRNA in cytoplasm Degradation of mRNA Translation Polypeptide Protein processing Active protein Degradation of protein Transport to cellular destination Cellular function Transcription 1 Promoter 5 3 3 5 TATA box DNA strand 2 Transcription factors 5 3 INITIATION 3 5 3 RNA polymerase 5 3 3 5 5 RNA transcript Transcription initiation complex Promoter Transcription unit 5 3 Start point RNA polymerase DNA 3 5 Promoter Transcription unit 5 3 Start point RNA polymerase 3 5 DNA 1 Initiation 5 3 Unwound DNA 3 5 RNA transcript Template strand of DNA Promoter Transcription unit 5 3 Start point RNA polymerase 3 5 DNA 1 Initiation 5 3 3 5 Unwound DNA RNA transcript Template strand of DNA 2 Elongation Rewound DNA 5 3 3 5 RNA transcript 3 5 Promoter Transcription unit 5 3 Start point RNA polymerase 3 5 DNA 1 Initiation 5 3 3 5 Unwound DNA RNA transcript Template strand of DNA 2 Elongation Rewound DNA 5 3 3 5 3 5 RNA transcript 3 Termination 5 3 3 5 5 Completed RNA transcript 3 Nontemplate strand of DNA Elongation RNA polymerase 3 RNA nucleotides 3 end 5 5 Direction of transcription (“downstream”) Newly made RNA Template strand of DNA NEXT?: After done making long hnRNA, what next? 1. Add the Poly-AAAAAA tail and GTP Cap (UTR = untranslated region) 5 G Protein-coding segment Polyadenylation signal 3 P P P 5 Cap 5 UTR Start codon AAUAAA Stop codon 3 UTR AAA…AAA Poly-A tail 2. (Eukaryotes) Remove Introns hn-RNA 5 Exon1 5 Cap Intron 1 30 Exon3 3 Exon2 40 Intron 100 Poly-A tail 120 146 SPLICING Coding segment mRNA 5 Cap 1 5 UTR Introns cut out and exons spliced together Poly-A tail 146 3 UTR Also Alternative Splicing can occur RNA transcript (hnRNA) 5 Exon 1 Protein snRNA Intron Exon 2 Other proteins snRNPs RNA transcript (hnRNA) 5 Exon 1 Intron Protein snRNA Exon 2 Other proteins snRNPs Spliceosome 5 RNA transcript (hnRNA) 5 Exon 1 Intron Protein snRNA Exon 2 Other proteins snRNPs Spliceosome 5 Spliceosome components 5 mRNA Exon 1 Exon 2 Cut-out Intron (lariat) The RNA Tie Club was right! RNA transcript (pre-mRNA) 5 Exon 1 Intron Protein snRNA Other proteins snRNPs Spliceosome snRNA -> Exon 2 5 3 Amino acid attachment site 5 tRNA -> Hydrogen bonds Anticodon (a) Two-dimensional structure SRP-RNA rRNA in ribosomes Ribosome mRNA Signal peptide Signal peptide removed Signalrecognition particle (SRP) Protein Insulin! CYTOSOL ER LUMEN ER membrane SRP receptor protein Translocation complex GFP = Green Florescent Protein (from Jelly fish) Activators Promoter DNA Enhancer Studying Promoters Distal control element TATA box GFP Gene Promoter Activators DNA Enhancer Distal control element GFP Gene TATA box General transcription factors DNA-bending protein Studying Promoters Group of mediator proteins Promoter Activators DNA Enhancer Distal control element GFP Gene TATA box General transcription factors DNA-bending protein Studying Promoters Group of mediator proteins RNA polymerase II RNA polymerase II Transcription initiation complex RNA synthesis GFP Enhancer Promoter Control elements GFP (blue) Albumin gene GFP (red) Crystallin gene LIVER CELL NUCLEUS LENS CELL NUCLEUS Available activators Available activators blue! Albumin gene expressed red! Crystallin gene not expressed (a) Liver cell Albumin gene not expressed Crystallin gene expressed (b) Lens cell “Brainbow” GFP= Nobel Prize 2008 Martin Chalfie, Roger Tsien and Osamu Shimomura DNA TRANSCRIPTION 3 5 RNA transcript RNA PROCESSING RNA polymerase Exon RNA transcript (pre-mRNA) Intron Aminoacyl-tRNA synthetase NUCLEUS Amino acid CYTOPLASM AMINO ACID ACTIVATION tRNA mRNA Growing polypeptide 3 A Activated amino acid P E Ribosomal subunits 5 TRANSLATION E A Codon Ribosome Anticodon
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