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Central Dogma DNA is the genetic material within the nucleus. Replication The process of replication creates new copies of DNA. The process of transcription creates an RNA using DNA information. DNA Transcription RNA Nucleus The process of translation creates a protein using RNA information. Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation Protein Cytoplasm Transcription • DNA is used as a template for creation of RNA using • the enzyme RNA polymerase. DNA 5’ G T C A T T C G G 3’ 3’ C A G T A A G C C Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 5’ Transcription • RNA polymerase reads the nucleotides on the • template strand from 3’ to 5’ and creates an RNA • Molecule in a 5’ to 3’ direction that looks like the codingGstrand. T C A T T C G G C A G T A A G C C Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Transcription • The new RNA molecule is formed by incorporating • nucleotides that are complementary to the template strand. DNA coding strand 5’ DNA G T C A T T C G G 3’ 3’ G U C A U U C G G 3’ C A G T A A G C C 5’ DNA template strand 5’ Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings RNA Two types of nucleic acids •RNA •DNA •Usually single-stranded •Usually double-stranded •Has uracil as a base •Has thymine as a base •Ribose as the sugar •Deoxyribose as the sugar •Carries protein-encoding information •Carries RNA-encoding information •Can be catalytic •Not catalytic Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Two types of nucleic acids # of strands kind of sugar bases used Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings rRNA is part of ribosome, used to translate mRNA into protein Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings tRNA is a connection between anticodon and amino acid Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Initiation of transcription Transcription begins at the 3’ end of the gene in a region called the promoter. The promoter recruits TATA protein, a DNA binding protein, which in turn recruits other proteins. TATA binding protein Promoter DNA GG Transcription factor Gene sequence to be transcribed TATA CCC TATA box Transcription begins When a complete transcription complex is formed RNA polymerase binds and transcription begins. Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 10.10 Eukaryotic RNA is processed before leaving the nucleus • Noncoding segments called introns are spliced out Exon Intron Exon Intron Exon DNA Cap RNA transcript with cap and tail • A cap and a tail are added to the ends to protect against degradation in the cytoplasm Transcription Addition of cap and tail Introns removed Tail Exons spliced together mRNA Coding sequence NUCLEUS CYTOPLASM Figure 10.10 Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Fig. 10.20 Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 10.8 The genetic code is the Rosetta stone of life • Virtually all organisms share the same genetic code • All organisms use the same 20 aa • Each codon specifies a particular aa Figure 10.8A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 10.8 The genetic code is the Rosetta stone of life • Three codons do not code from an aa • Rather they are found at the end of the coding sequence • Tell a ribosome to stop translation and release the protein Figure 10.8A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings • Tryptophan and Methionine have only 1 codon each • All the rest have more than one • AUG has a dual function • 3 stop codons that code for termination of protein synthesis • Redundancy in the code but no ambiguity Figure 10.8A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation • The process of reading the RNA sequence of an mRNA and creating the amino acid sequence of a protein is called translation. DNA Transcription T T C A G T C A G A A G U C A G U C DNA template strand Messenger RNA mRNA Codon Codon Codon Translation Protein Lysine Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Serine Valine Polypeptide (amino acid sequence) 10.11 Transfer RNA molecules serve as interpreters during translation • In the cytoplasm, a ribosome attaches to the mRNA and translates its message into a polypeptide • The process is aided by transfer RNAs Amino acid attachment site Hydrogen bond RNA polynucleotide chain Anticodon Figure 10.11A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings A codon of three nucleotides determines choice of amino acid Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation is composed of three steps • Initiation translation begins at start codon (AUG=methionine) Elongation the ribosome uses the tRNA anticodon to match codons to amino acids and adds those amino acids to the growing peptide chain Termination translation ends at the stop codon UAA, UAG or UGA Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings • mRNA, a specific tRNA, and the ribosome subunits assemble during initiation Large Ribosomal subunit Initiator tRNA P site A site Start codon mRNA binding site Small ribosomal subunit 1 Figure 10.13B Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 2 Translation initiation Leader sequence Small ribosomal subunit 5’ 3’ mRNA mRNA U U C G U C A U G G G A U G U A A G C G A A U A C Assembling to begin translation Initiator tRNA Met Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation Elongation Ribosome 5’ 3’ mRNA A U G G G A U G U A A G C G A U A C C C U tRNA P A Amino acid Met Gly Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Large ribosomal subunit Translation Elongation 5’ 3’ mRNA A U G G G A U G U A A G C G A U A C C C U A A C A P Met Gly Cys Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation Elongation 5’ 3’ mRNA A U G G G A U G U A A G C G A P C C U A C A U U C Cys Gly Lengthening polypeptide (amino acid chain) Me t Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Lys A Translation Elongation Stop codon 5’ mRNA A U G G G A U G U A A G C G A U A A C U U C G C U A A A P Cy s Gly et M Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Lys Arg Release factor Translation Termination Stop codon Ribosome reaches stop codon 5’ mRNA A U G G G A U G U A A G C G A U A A G C U U P U C Release factor Arg Lys Met Gly Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Cys A Translation Termination Once stop codon is reached, elements disassemble. A U G G GA UG U AA G C G A U A A G C P U Release factor A rg s Cy Met Gly Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings s Ly A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Levels of protein structure Primary structure Secondary structure Tertiary structure sequence of amino acids shapes formed with regions of the protein (helices, coil, sheets) shape of entire folded protein due to interactions between particular peptides Quaternary structure structures formed by interaction of several proteins together e.g. Functional hemoglobin is two alpha-hemoglobin proteins and two beta-hemoglobin proteins Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 10_14d.jpg Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Levels of protein structure Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Misfolding of protein impairs function •Misfolded prion protein disrupts functions of other normally folded prion proteins. •Aberrant conformation can passed on propagating like an “infectious” agent. Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 10_18.jpg Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings