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Using DNA -Transcription - RNA processing - Universality of the genetic code - Translation Refer to chapter 12.3 to 12.6, and 13.2, 13.3 in text. There are enzymes that can make proteins (nonribosomal peptides) without this process: Those are enzyme-specific, and not what we are talking about here. Review DNA structure (What does it look like?) and RNA structure. Recap DNA replication (Unzipping the... helix, etc.) So... what do you recall/know about transcription and translation? purpose? location? general mechanisms? Now go to animations. TRANSCRIPTION/mRNA PROCESSING Transcription and processing only https://www.youtube.com/watch?v=LPnz1i0Fr98 Also to processing only https://www.youtube.com/watch?v=eMYl0wjNc0U ‘realistic” https://www.youtube.com/watch?v=5MfSYnItYvg You can also go to Crash Course or Bozeman of course transcription: DNA to RNA … that promoter thing isn’t as simple as the previous picture suggests! http://www.google.com/imgres?imgurl=http://biology.kenyon.edu/courses/biol114/Chap10 (You don’t need to worry about this. I just don’t want you to think you are responsible for everything! There will still be stuff to learn in college!) processing of pre-mRNA: 5’ cap, poly(A) tail ↑, and excision of introns →, leaving exons to move out of the nucleus… Some pre-mRNA are processed to include different exons One gene can code for multiple proteins. Some introns become microRNAs (miRNA) that impact translation… Are you accessing the IB syllabus at the Stanton web site? Transcription and RNA processing terminology (Read all as bold and underlined.) initiation promoter transcription factor elongation 5’ to 3’ RNA polymerase transcription unit termination terminator termination point sense strand antisense (template) strand pre-mRNA leader trailer intron exon snRNPs spliceosome 5’ cap poly A tail mRNA So now the message can leave the engineering library and go out onto the factory floor. The ‘factory’ has 20 amino acids, while the plans only have 4 nucleotides.... Genetic Code Deciphered by Marshall Nirenberg, 1966. Note - how table is read. - what it represents (RNA codons). - start codon - stop codons - redundancy: most amino acids have multiple codons… (ramifications?) What amino acid does UGU code for? What is the codon for alanine? What is a DNA sequence for met-ser-his? What a.a. sequence from TCAGGAACAATTTAG? Same code, different presentation (Some tables have the DNA rather than the RNA sequences.) Code is almost universal - exceptions usually mitochondrial - often involve a.a. for stop codon(s) - may swap in more stops - novel amino acids: usually in prokaryotes pyrrolysine for UAG selenocysteine for some UGA (25 of your proteins have this last one!) http://www.biology.lsu.edu/heydrjay/1201/Chapter17/SCI_Amino_Acid_CIRCLE.jpg go to an animation for translation… TRANSLATION “Basic”, but fairly complete… https://www.youtube.com/watch?v=8dsTvBaUMvw Virtual Cell https://www.youtube.com/watch?v=5bLEDd-PSTQ Realistic view https://www.youtube.com/watch?v=TfYf_rPWUdY translation: from the language of nucleic acids, into that of protein. nobelprize.org/.../medicine/dna/a/translation/ ↑ NB close-up of tRNA → aminoacyl-tRNA synthetase ↓ ←NB www.nsf.gov/news/overviews/biology/interact05.jsp This is happening on rER ↑ if the proteins are for export or are lysozymes; Local proteins are made on free ribosomes. The shift from the A to P site is translocation. Recall the peptide bond… and be able to draw it. → The energy required for the formation of one peptide bond includes the hydrolysis of 2 ATP and 2 GTP molecules. Regulation of gene expression in eukaryotes chromatin structure methylation/acetylation of histone tails, epigenetics, inherited factors transcriptional control methylation of cytosines blocks reading role in cell specialization see also slide 4 posttranscriptional control alternate splicing control of mRNA exiting nucleus translational control persistence of 5’ cap and poly-A tail microRNA from introns → RISC (RNA-induced silencing complex) posttranslational control tertiary structure, prosthetic groups? coenzymes? proteases? lifted from p. 237 in Mader Different genes coordinate whole structures Pax6 (eyes), Tbx5 (limbs), Hox (body shape) Translation terminology (Read all as bold and underlined.) initiation mRNA codon start codon tRNA amino acid CCA tail (amino acid binding site) tRNA-activating enzyme (aminoacyl-tRNA synthetase) anticodon ribosome rRNA ribosomal protein nucleolus small ribosomal subunit mRNA binding site elongation large ribosomal subunit A, P and E sites translocation 5’ to 3’ (reverse relative to DNA) polysome termination stop codon What is meant by the genetic code? How is it “universal”? What are the three stages of DNA to protein? Where does each occur? Walk through each. There were 4 different RNAs involved: Identify each along with its use. Give examples of form fitting function, for ribosomes and tRNA. Lots of proteins… Investigate any for which the name does not obviously lead to the job. transcription intron CCA tail initiation exon amino acid binding site promoter snRPs tRNA-activating enzyme transcription factor spliceosome aminoacyl-tRNA synthetase elongation 5’ cap anticodon 5’ to 3’ poly A tail ribosome RNA polymerase mRNA rRNA transcription unit start codon ribosomal protein termination nucleolus terminator (Arnold) redundancy small ribosomal subunit termination point The Genetic Code mRNA binding site sense strand translation elongation antisense strand translocation large ribosomal subunit pre-mRNA initiation A, P and E sites leader codon polysome trailer tRNA termination amino acid