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BIOLOGY CONCEPTS & CONNECTIONS Fourth Edition Neil A. Campbell • Jane B. Reece • Lawrence G. Mitchell • Martha R. Taylor CHAPTER 10 Protein Synthesis From PowerPoint® Lectures for Biology: Concepts & Connections Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings THE FLOW OF GENETIC INFORMATION FROM DNA TO RNA TO PROTEIN The DNA genotype is expressed as proteins, which provide the molecular basis for phenotypic traits • The information constituting an organism’s genotype is carried in the sequence of bases in DNA • The flow of information is from DNA to RNA to protein Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings • A specific gene specifies a polypeptide – The DNA is transcribed into RNA, which is translated into the polypeptide http://www.wiley.com/legacy/college/boyer/0470003790/animations/central_dogma/central_dogma.swf DNA TRANSCRIPTION RNA TRANSLATION Protein Figure 10.6A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings • Studies of inherited metabolic disorders first suggested that phenotype is expressed through proteins • Studies of the bread mold Neurospora crassa led to the one gene-one polypeptide hypothesis Figure 10.6B Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Mutate wild type fungus *Supply all mutant isolates with complete media *Grow purified mutants with minimal media to find nutritional mutants *Determine what is the nutritional limitation find mutation Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings There for the gene used to produce an enzyme that helps cells manufacture Arginine amino acid was mutated in that fungal strain Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Transcription produces genetic messages in the form of RNA RNA polymerase RNA nucleotide Direction of transcription Template strand of DNA Figure 10.9A Newly made RNA Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings RNA Transcription • Process in which the genetic information on DNA is transferred to RNA • During transcription only 1 DNA stand serves as the template or pattern from which RNA is formed. Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings RNA polymerase • In transcription, the DNA helix unzips – RNA nucleotides line up along one strand of the DNA following the base-pairing rules – The single-stranded messenger RNA peels away and the DNA strands rejoin DNA of gene Promoter DNA Initiation Elongation Terminator DNA Area shown in Figure 10.9A Termination Growing RNA Completed RNA http://www.stolaf.edu/people/giannini/flashanimat/molgenetics/transcription.swf Figure 10.9B Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings RNA polymerase RNA Transcription 1. Initiation • The enzyme RNA polymerase attaches to the promoter site on the DNA • Promoter – a sequence of nucleotides that is found on one of the DNA strands – tells RNA polymerase to start transcription and which of the two DNA strands to transcribe Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings RNA Transcription 2. Elongation • RNA nucleotides attach to the free DNA nucleotides by hydrogen bonds one at a time • As RNA synthesis continues the growing RNA strand peels away from the DNA and the DNA strands rejoin Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings RNA Transcription 3. Termination • RNA polymerase reaches the terminator. • Terminator – a sequence of bases on DNA that signals the end of the gene • The RNA polymerase detaches from the DNA and the RNA molecule is complete Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 10.10 Eukaryotic RNA is processed before leaving http://www.four-h.purdue.edu/apple_genomics/flash/movie3.swf the nucleus http://highered.mcgraw-hill.com/sites/0072556781/student_view0/chapter14/animation_quiz_3.html • Noncoding segments called introns are spliced out Exon Intron Intron Exon DNA Cap RNA transcript with cap and tail • The coding segments called exons are joined together • A cap and a tail are added to the ends Exon 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 Genetic information written in codons is translated into amino acid sequences • The “words” of the DNA “language” are triplets of bases called codons – The codons in a gene specify the amino acid sequence of a polypeptide Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Gene 1 Gene 3 DNA molecule Gene 2 DNA strand TRANSCRIPTION RNA Codon TRANSLATION Polypeptide Figure 10.7 Amino acid Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings The genetic code is the Rosetta stone of life • Virtually all organisms share the same genetic code Figure 10.8A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings • An exercise in translating the genetic code Transcribed strand DNA Transcription RNA Start codon Polypeptide Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation Stop codon Figure 10.8B Translation • The process in which a polypeptide is synthesized using the genetic information encoded on an mRNA molecule • The following are needed for translation to occur 1. mRNA - Contains the instructions for the assembly of proteins - Codon – a sequence of 3 bases on mRNA that specifies a specific amino acid that will be added to the polypeptide chain Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 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 • Each tRNA molecule has a triplet anticodon on one end and an amino acid attachment site on the other Amino acid attachment site Anticodon Figure 10.11B, C Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation 2. tRNA (transfer RNA) • Carries an amino acid to the ribosome • A tRNA molecule is composed of – A single strand of RNA (about 80 nucleotides) – A loop at one end that contains the anticodon – Anticodon – a sequence of 3 bases on tRNA that are complementary to the bases on mRNA – At the opposite end of the loop is a site where an amino acit can attach Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation 3. Amino acids • Located in the cytoplasm • Synthesized from other chemicals or obtained from food Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 10.12 Ribosomes build polypeptides Next amino acid to be added to polypeptide Growing polypeptide tRNA molecules P site A site Growing polypeptide Large subunit tRNA P A mRNA mRNA binding site Codons mRNA Small subunit Figure 10.12A-C Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Translation 4. Ribosomes • Organelles where protein synthesis occurs • Consists of 2 subunits each made up of proteins and ribosomal RNA (rRNA) – Small subunit – has binding site for mRNA – Large subunit – has binding site for tRNA Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings An initiation codon marks the start of an mRNA message Start of genetic message End Figure 10.13A 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 Small ribosomal subunit 1 Figure 10.13B Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 2 New peptide bond forming Growing polypeptide Codons Stage 4 Elongation A succession of tRNAs add their amino acids to the polypeptide chain as the mRNA is moved through the ribosome, one codon at a time. mRNA Polypeptide Stop Codon Figure 10.15 (continued) Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Stage 5 Termination The ribosome recognizes a stop codon. The polypeptide is terminated and released. 10.14 Elongation adds amino acids to the polypeptide chain until a stop codon terminates translation • The mRNA moves a codon at a time relative to the ribosome – A tRNA pairs with each codon, adding an amino acid to the growing polypeptide Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Amino acid Polypeptide A site P site Anticodon mRNA 1 Codon recognition mRNA movement Stop codon New peptide bond 3 Translocation Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings 2 Peptide bond formation Figure 10.14 Steps of Translation 1. Initiation • mRNA binds to the ribosome • The start codon (AUG) is reached • The first amino acid (methionine) is brought to the ribosome by the tRNA 2. Elongation • Amino acids are added one by one to a growing polypeptide chain Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Steps of Translation 3. Termination • The stop codon is reached • The completed polypeptide is released Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Modification of the polypeptide Endoplasmic reticulum • Collects proteins made by the ribosomes • Packages them into vesicles which move to the Golgi apparatus Golgi apparatus • Proteins are altered, packaged into vesicles, and transported to different parts of the cell or exported out of the cell Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings • Summary of transcription and translation TRANSCRIPTION DNA mRNA RNA polymerase Stage 1 mRNA is transcribed from a DNA template. Amino acid TRANSLATION Enzyme Stage 2 Each amino acid attaches to its proper tRNA with the help of a specific enzyme and ATP. tRNA Initiator tRNA mRNA Figure 10.15 Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Anticodon Large ribosomal subunit Start Codon Small ribosomal subunit Stage 3 Initiation of polypeptide synthesis The mRNA, the first tRNA, and the ribosomal subunits come together. Review: The flow of genetic information in the cell is DNARNAprotein • The sequence of codons in DNA spells out the primary structure of a polypeptide – Polypeptides form proteins that cells and organisms use Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Mutations can change the meaning of genes • Mutations are changes in the DNA base sequence – These are caused by errors in DNA replication or by mutagens – The change of a single DNA nucleotide causes sickle-cell disease Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Normal hemoglobin DNA mRNA Mutant hemoglobin DNA mRNA Normal hemoglobin Sickle-cell hemoglobin Glu Val http://www.cleanvideosearch.com/media/action/yt/watch?v=1fN7rOwDyMQ&safety_mode=true&persist_safety_mode=1&safe=active Figure 10.16A Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings • Types of mutations NORMAL GENE mRNA Protein Met Lys Phe Gly Ala Lys Phe Ser Ala BASE SUBSTITUTION Met Missing BASE DELETION Met Lys Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Leu Ala His Figure 10.16B Types of Mutations There are 2 general categories of mutations: 1. Base substitution • The replacement of one nucleotide with another • Can result in no change in the protein • An insignificant change – The altered amino acid has no effect on the function of the protein Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Types of Mutations • A change that is crucial to life of the organism – The altered amino acid has an effect on the function of the protein 2. Base insertions or deletions • One or more bases are added or deleted from the DNA • Often have disastrous effects – The nucleotide sequence following the change alters the genetic message (reading frame) Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings Mutations are Useful Mutations are useful because they 1. Provide diversity that allows evolution by natural selection to occur 2. Essential tool for geneticists • Create different alleles needed for genetic research Copyright © 2003 Pearson Education, Inc. publishing as Benjamin Cummings