From Gene to Protein. Chapter 17. Biology Eighth Edition Neil Campbell and Jane Reece. PowerPoint Lecture Presentations for

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1 Chapter 17 From Gene to Protein PowerPoint Lecture Presentations for Biology Eighth Edition Neil Campbell and Jane Reece Lectures by Chris Romero, updated by Erin Barley with contributions from Joan Sharp Copyright 2008 Pearson Education, Inc., publishing as Pearson Benjamin Cummings

2 Overview: The Flow of Genetic Information The information content of DNA is in the form of specific sequences of nucleotides The DNA inherited by an organism leads to specific traits by dictating the synthesis of proteins Proteins are the links between genotype and phenotype Gene expression, the process by which DNA directs protein synthesis, includes two stages: transcription and translation 2

3 Genes specify proteins via transcription and translation How was the fundamental relationship between genes and proteins discovered? 3

4 Nutritional Mutants in Neurospora: Scientific Inquiry George Beadle and Edward Tatum exposed bread mold to X-rays, creating mutants that were unable to survive on minimal medium as a result of inability to synthesize certain molecules They developed a one gene one enzyme hypothesis, which states that each gene dictates production of a specific enzyme 4

5 Condition EXPERIMENT Growth: Wild-type cells growing and dividing No growth: Mutant cells cannot grow and divide Minimal medium RESULTS Minimal medium (MM) (control) Wild type Classes of Neurospora crassa Class I mutants Class II mutants Class III mutants MM + ornithine MM + citrulline MM + arginine (control) CONCLUSION Wild type Class I mutants (mutation in gene A) Class II mutants (mutation in gene B) Class III mutants (mutation in gene C) Gene A Gene B Gene C Precursor Precursor Precursor Precursor Enzyme A Enzyme A Enzyme A Enzyme A Ornithine Ornithine Ornithine Ornithine Enzyme B Enzyme B Enzyme B Enzyme B Citrulline Citrulline Citrulline Citrulline Enzyme C Enzyme C Enzyme C Enzyme C Arginine Arginine Arginine Arginine

6 The Products of Gene Expression: A Developing Story Some proteins aren t enzymes, so researchers later revised the hypothesis: one gene one protein Many proteins are composed of several polypeptides, each of which has its own gene Therefore, Beadle and Tatum s hypothesis is now restated as the one gene one polypeptide hypothesis Note that it is common to refer to gene products as proteins rather than polypeptides 6

7 Basic Principles of Transcription and Translation RNA is the intermediate between genes and the proteins for which they code Transcription is the synthesis of mrna under the direction of DNA Transcription produces messenger RNA (mrna) Translation is the synthesis of a polypeptide, which occurs under the direction of mrna Ribosomes are the sites of translation 7

8 In prokaryotes, mrna produced by transcription is immediately translated without more processing In a eukaryotic cell, the nuclear envelope separates transcription from translation The central dogma is the concept that cells are governed by a cellular chain of command (the flow of information): DNA RNA protein 8

9 TRANSCRIPTION DNA Overview: the roles of transcription and translation in the flow of genetic information TRANSLATION Polypeptide mrna Ribosome (a) Bacterial cell Nuclear envelope TRANSCRIPTION DNA RNA PROCESSING Pre-mRNA mrna TRANSLATION Ribosome Polypeptide (b) Eukaryotic cell

10 Codons: Triplets of Bases The flow of information from gene to protein is based on a triplet code: a series of nonoverlapping, three-nucleotide words These triplets are the smallest units that can code for all the amino acids 10

11 During transcription, one of the two DNA strands called the template strand provides a template for ordering the sequence of nucleotides in an RNA transcript During translation, the mrna base triplets, called codons, are read in the to direction Each codon specifies the amino acid to be placed at the corresponding position along a polypeptide 11

12 DNA molecule Gene 1 Gene 2 Gene 3 The triplet code DNA template strand TRANSCRIPTION mrna Codon TRANSLATION Protein Amino acid

13 First mrna base ( end of codon) Third mrna base ( end of codon) Of the 64 triplets, 61 code for amino acids; 3 triplets are stop signals to end translation The genetic code is redundant but not ambiguous; no codon specifies more than one amino acid Second mrna base

14 Evolution of the Genetic Code The genetic code is nearly universal, shared by the simplest bacteria to the most complex animals Genes can be transcribed and translated after being transplanted from one species to another 14

15 Expression of genes from different species (a) Tobacco plant expressing a firefly gene (b) Pig expressing a jellyfish gene

16 Molecular Components of Transcription Transcription, the first stage of gene expression, can be examined in more detail RNA synthesis is catalyzed by RNA polymerase, which pries the DNA strands apart and hooks together the RNA nucleotides RNA synthesis follows the same basepairing rules as DNA, except uracil substitutes for thymine 16

17 RNA polymerase RNA nucleotides Direction of transcription Newly made RNA Template strand of DNA 17

18 The DNA sequence where RNA polymerase attaches is called the promoter; in bacteria, the sequence signaling the end of transcription is called the terminator The stretch of DNA that is transcribed is called a transcription unit Animation: Transcription 18

19 Synthesis of an RNA Transcript The three stages of transcription: Initiation Elongation Termination 19

20 Promoters signal the initiation of RNA synthesis Elongation: as RNA polymerase moves along the DNA, it untwists the double helix Transcription progresses at a rate of 40 nucleotides per second in eukaryotes A gene can be transcribed simultaneously by several RNA polymerases Termination is the end of mrna synthesis 20

21 Figure Promoter Transcription unit Start point DNA RNA polymerase 1 Initiation Unwound DNA Nontemplate strand of DNA Template strand of DNA RNA transcript 2 Elongation Rewound DNA RNA transcript 3 Termination Completed RNA transcript Direction of transcription ( downstream ) 21

22 Each end of a pre-mrna molecule is modified in a particular way: The end receives a modified nucleotide cap The end gets a poly-a tail These modifications are mainly to protect mrna from hydrolytic enzymes G P Protein-coding segment Polyadenylation signal P P AAUAAA AAA AAA Cap UTR Start codon Stop codon UTR Poly-A tail 22

23 Split Genes and RNA Splicing Most eukaryotic genes and their RNA transcripts have long noncoding stretches of nucleotides that lie between coding regions These noncoding regions are called intervening sequences, or introns The other regions are called exons because they are eventually expressed, usually translated into amino acid sequences RNA splicing removes introns and joins exons, creating an mrna molecule with a continuous coding sequence 23

24 Ribozymes Ribozymes are catalytic RNA molecules that function as enzymes and can splice RNA The discovery of ribozymes rendered obsolete the belief that all biological catalysts were proteins 2011 Pearson Education, Inc.

25 RNA processing: RNA splicing Pre-mRNA Cap Exon Intron Exon Intron 105 Exon 146 Poly-A tail Coding segment Introns cut out and exons spliced together mrna Cap UTR UTR Poly-A tail

26 The Functional and Evolutionary Importance of Introns Some introns contain sequences that may regulate gene expression Some genes can encode more than one kind of polypeptide, depending on which segments are treated as exons during splicing This is called alternative RNA splicing Consequently, the number of different proteins an organism can produce is much greater than its number of genes 2011 Pearson Education, Inc.

27 Translation is the RNA-directed synthesis of a polypeptide: a closer look The translation of mrna to protein can be examined in more detail 27

28 Molecular Components of Translation A cell translates an mrna message into protein with the help of transfer RNA (trna) Molecules of trna are not identical: Each carries a specific amino acid on one end Each has an anticodon on the other end; the anticodon base-pairs with a complementary codon on mrna BioFlix: Protein Synthesis 28

29 Translation: the basic concept Polypeptide Amino acids Ribosome trna with amino acid attached trna Anticodon mrna Codons

30 Flattened into one plane to reveal its base pairing, a trna molecule looks like a cloverleaf Because of hydrogen bonds, trna actually twists and folds into a threedimensional molecule Amino acid attachment site Anticodon (a) Two-dimensional structure Anticodon Hydrogen bonds Hydrogen bonds (b) Three-dimensional structure Amino acid attachment site Anticodon (c) Symbol used in this book A trna molecule consists of a single RNA strand that is short - only about 80 nucleotides long

31 Accurate translation requires two steps: First: a correct match between a trna and an amino acid Second: a correct match between the trna anticodon and an mrna codon Ribosomes Ribosomes facilitate specific coupling of trna anticodons with mrna codons in protein synthesis The two ribosomal subunits (large and small) are made of proteins and ribosomal RNA (rrna) 31

32 The anatomy of a functioning ribosome trna molecules Growing polypeptide E P A Exit tunnel Large subunit Small subunit mrna (a) Computer model of functioning ribosome P site (Peptidyl-tRNA binding site) E site (Exit site) mrna binding site E P A A site (AminoacyltRNA binding site) Large subunit Small subunit (b) Schematic model showing binding sites Amino end mrna E Growing polypeptide Next amino acid to be added to polypeptide chain trna Codons (c) Schematic model with mrna and trna

33 Figure 17.17a trna molecules Growing polypeptide Exit tunnel E P A Large subunit Small subunit mrna (a) Computer model of functioning ribosome

34 A ribosome has three (in some sources two) binding sites for trna: The P site holds the trna that carries the growing polypeptide chain The A site holds the trna that carries the next amino acid to be added to the chain [The E site*] is the exit site, where discharged trnas leave the ribosome * many molecular biologists do NOT consider that as a binding site 34

35 Building a Polypeptide The three stages of translation are similar to those of transcription: Initiation Elongation Termination 35

36 Ribosome Association and Initiation of Translation The initiation stage of translation brings together mrna, a trna with the first amino acid, and the two ribosomal subunits First, a small ribosomal subunit binds with mrna and a special initiator trna Then the small subunit moves along the mrna until it reaches the start codon (AUG) Proteins called initiation factors bring in the large subunit that completes the translation initiation complex 36

37 The initiation of translation U A A U C G P site Large ribosomal subunit Initiator trna mrna Start codon mrna binding site Small ribosomal subunit GTP GDP E A Translation initiation complex

38 Elongation of the Polypeptide Chain During the elongation stage, amino acids are added one by one to the preceding amino acid Each addition occurs in three steps: codon recognition, peptide bond formation, and translocation 38

39 Amino end of polypeptide mrna E Ribosome ready for next aminoacyl trna P site A site GTP GDP E E P A P A The elongation cycle of translation GDP GTP E P A

40 Figure 17.17a trna molecules Growing polypeptide Exit tunnel E P A Large subunit Small subunit mrna (a) Computer model of functioning ribosome

41 Termination of Translation Termination occurs when a stop codon in the mrna reaches the A site of the ribosome The A site accepts a protein called a release factor The release factor causes the addition of a water molecule instead of an amino acid This reaction releases the polypeptide, and the translation assembly then comes apart 41

42 The termination of translation Release factor Free polypeptide Stop codon (UAG, UAA, or UGA) 2 GTP 2 GDP

43 Polyribosomes A number of ribosomes can translate a single mrna simultaneously, forming a polyribosome (or polysome) Polyribosomes enable a cell to make many copies of a polypeptide very quickly Incoming ribosomal subunits (a) Growing polypeptides Start of mrna ( end) End of mrna ( end) Ribosomes mrna Completed polypeptide (b) 0.1 µm

44 Protein Folding and Post-Translational Modifications During and after synthesis, a polypeptide chain spontaneously coils and folds into its three-dimensional shape Polypeptides may come together to form the subunits of a protein 44

45 Targeting Polypeptides to Specific Locations Two populations of ribosomes are evident in cells: free ribsomes (in the cytosol) and bound ribosomes (attached to the ER) Free ribosomes mostly synthesize proteins that function in the cytosol Bound ribosomes make proteins of the endomembrane system and proteins that are secreted from the cell Ribosomes are identical and can switch from free to bound 45

46 DNA Transcription RNA Nucleus Cytoplasm Translation Protein DNA RNA protein Flow of genetic information in a eukaryotic cell 46

47 Review: The flow of genetic information in the cell is DNA RNA protein Does translation represent: DNA RNA or RNA protein? Where does the information for producing a protein originate: DNA or RNA? Which one has a linear sequence of codons: rrna, mrna, or trna? Which one directly influences the phenotype: DNA, RNA, or protein? 47 Copyright 2009 Pearson Education, Inc.

48 Point mutations can affect protein structure and function Mutations are changes in the genetic material of a cell or virus Point mutations are chemical changes in just one base pair of a gene The change of a single nucleotide in a DNA template strand can lead to the production of an abnormal protein 48

49 Wild-type hemoglobin DNA C T T G A A Mutant hemoglobin DNA C A T G T A mrna G A A mrna G U A Normal hemoglobin Glu Sickle-cell hemoglobin Val

50 Types of Point Mutations Point mutations within a gene can be divided into two general categories: Base-pair substitutions Base-pair insertions or deletions 50

51 Types of point mutations DNA template strand mrna Protein Amino end A instead of G Wild-type Extra A Stop Carboxyl end U instead of C Extra U Stop Silent (no effect on amino acid sequence) Stop Frameshift causing immediate nonsense (1 base-pair insertion) T instead of C missing A instead of G missing Stop Missense Frameshift causing extensive missense (1 base-pair deletion) A instead of T missing U instead of A missing Nonsense Stop Stop No frameshift, but one amino acid missing (3 base-pair deletion) (a) Base-pair substitution (b) Base-pair insertion or deletion

52 Substitutions A base-pair substitution replaces one nucleotide and its partner with another pair of nucleotides Silent mutations have no effect on the amino acid produced by a codon because of redundancy in the genetic code Missense mutations still code for an amino acid, but not necessarily the right amino acid Nonsense mutations change an amino acid codon into a stop codon, nearly always leading to a nonfunctional protein 52

53 Insertions and Deletions Insertions and deletions are additions or losses of nucleotide pairs in a gene These mutations have a disastrous effect on the resulting protein more often than substitutions do Insertion or deletion of nucleotides may alter the reading frame, producing a frameshift mutation 53

54 Mutagens Mutagens are physical or chemical agents that can cause mutations High-energy radiation Chemicals Etc. 54

55 Comparing Gene Expression in Bacteria, Archaea, and Eukarya Bacteria and Eukarya differ in their RNA polymerases, termination of transcription and ribosomes; Archaea tend to resemble Eukarya in these respects Bacteria can simultaneously transcribe and translate the same gene In Eukarya, transcription and translation are separated by the nuclear envelope In Archaea, transcription and translation are likely coupled 55

56 What Is a Gene? Revisiting the Question The idea of the gene itself is a unifying concept of life We have considered a gene as: A discrete unit of inheritance A region of specific nucleotide sequence in a chromosome A DNA sequence that codes for a specific polypeptide chain 56

57 Figure RNA polymerase Coupled transcription and translation in bacteria RNA polymerase DNA Polyribosome Direction of transcription mrna 0.25 m DNA Polyribosome Polypeptide (amino end) Ribosome mrna ( end) 57

58 TRANSCRIPTION DNA A summary of transcription and translation in a eukaryotic cell RNA transcript RNA PROCESSING CYTOPLASM E A P Exon NUCLEUS Ribosomal subunits mrna RNA polymerase RNA transcript (pre-mrna) Intron Growing polypeptide Amino acid trna Aminoacyl-tRNA synthetase Activated amino acid AMINO ACID ACTIVATION TRANSLATION E A Anticodon Codon Ribosome

59 Figure 17.UN06d

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