Chapter 11 How Genes Are Controlled

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1 Chapter 11 How Genes Are Controlled PowerPoint Lectures for Biology: Concepts & Connections, Sixth Edition Campbell, Reece, Taylor, Simon, and Dickey Copyright 2009 Pearson Education, Inc. Lecture by Mary C. Colavito

2 Introduction: Cloning to the Rescue? Cloning has been attempted to save endangered species A clone is produced by asexual reproduction and is genetically identical to its parent Dolly the sheep was the first cloned mammal Endangered animals that were cloned include cows, oxen, sheep, wildcats, and wolves Disadvantages of cloning Does not increase genetic diversity Cloned animals may have health problems related to abnormal gene regulation Copyright 2009 Pearson Education, Inc.

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6 CONTROL OF GENE EXPRESSION Copyright 2009 Pearson Education, Inc.

7 11.1 Proteins interacting with DNA turn prokaryotic genes on or off in response to environmental changes Gene expression is the overall process of information flow from genes to proteins Mainly controlled at the level of transcription A gene that is turned on is being transcribed to produce mrna that is translated to make its corresponding protein Organisms respond to environmental changes by controlling gene expression Copyright 2009 Pearson Education, Inc.

8 11.1 Proteins interacting with DNA turn prokaryotic genes on or off in response to environmental changes An operon is a group of genes under coordinated control in bacteria The lactose (lac) operon includes Three adjacent genes for lactose-utilization enzymes Promoter sequence where RNA polymerase binds Operator sequence is where a repressor can bind and block RNA polymerase action Copyright 2009 Pearson Education, Inc.

9 11.1 Proteins interacting with DNA turn prokaryotic genes on or off in response to environmental changes Regulation of the lac operon Regulatory gene codes for a repressor protein In the absence of lactose, the repressor binds to the operator and prevents RNA polymerase action Lactose inactivates the repressor, so the operator is unblocked Copyright 2009 Pearson Education, Inc.

10 11.1 Proteins interacting with DNA turn prokaryotic genes on or off in response to environmental changes Types of operon control Inducible operon (lac operon) Active repressor binds to the operator Inducer (lactose) binds to and inactivates the repressor Repressible operon (trp operon) Repressor is initially inactive Corepressor (tryptophan) binds to the repressor and makes it active For many operons, activators enhance RNA polymerase binding to the promoter Copyright 2009 Pearson Education, Inc.

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12 Regulatory gene Promoter Operator OPERON Lactose-utilization genes DNA mrna Protein Active repressor RNA polymerase cannot attach to promoter Operon turned off (lactose absent) DNA mrna RNA polymerase bound to promoter Protein Lactose Inactive repressor Operon turned on (lactose inactivates repressor) Enzymes for lactose utilization

13 Regulatory gene Promoter Operator OPERON Lactose-utilization genes DNA mrna Protein Active repressor RNA polymerase cannot attach to promoter Operon turned off (lactose absent)

14 DNA mrna RNA polymerase bound to promoter Protein Lactose Inactive repressor Operon turned on (lactose inactivates repressor) Enzymes for lactose utilization

15 DNA Promoter Operator Gene Active repressor Active repressor Tryptophan Inactive repressor Lactose Inactive repressor lac operon trp operon

16 11.2 Differentiation results from the expression of different combinations of genes Differentiation involves cell specialization, in both structure and function Differentiation is controlled by turning specific sets of genes on or off Copyright 2009 Pearson Education, Inc.

17 Muscle cell Pancreas cells Blood cells

18 Muscle cell

19 Pancreas cells

20 Blood cells

21 11.3 DNA packing in eukaryotic chromosomes helps regulate gene expression Eukaryotic chromosomes undergo multiple levels of folding and coiling, called DNA packing Nucleosomes are formed when DNA is wrapped around histone proteins Beads on a string appearance Each bead includes DNA plus 8 histone molecules String is the linker DNA that connects nucleosomes Tight helical fiber is a coiling of the nucleosome string Supercoil is a coiling of the tight helical fiber Metaphase chromosome represents the highest level of packing DNA packing can prevent transcription Copyright 2009 Pearson Education, Inc.

22 11.3 DNA packing in eukaryotic chromosomes helps regulate gene expression Animation: DNA Packing Copyright 2009 Pearson Education, Inc.

23 DNA double helix (2-nm diameter) Linker Metaphase chromosome Tight helical fiber (30-nm diameter) Beads on a string Histones Nucleosome (10-nm diameter) Supercoil (300-nm diameter) 700 nm

24 11.4 In female mammals, one X chromosome is inactive in each somatic cell X-chromosome inactivation In female mammals, one of the two X chromosomes is highly compacted and transcriptionally inactive Random inactivation of either the maternal or paternal chromosome Occurs early in embryonic development and all cellular descendants have the same inactivated chromosome Inactivated X chromosome is called a Barr body Tortoiseshell fur coloration is due to inactivation of X chromosomes in heterozygous female cats Copyright 2009 Pearson Education, Inc.

25 Early embryo Two cell populations in adult X chromosomes Cell division and random X chromosome inactivation Active X Inactive X Orange fur Allele for orange fur Allele for black fur Inactive X Active X Black fur

26 11.5 Complex assemblies of proteins control eukaryotic transcription Eukaryotic genes Each gene has its own promoter and terminator Are usually switched off and require activators to be turned on Are controlled by interactions between numerous regulatory proteins and control sequences Copyright 2009 Pearson Education, Inc.

27 11.5 Complex assemblies of proteins control eukaryotic transcription Regulatory proteins that bind to control sequences Transcription factors promote RNA polymerase binding to the promoter Activator proteins bind to DNA enhancers and interact with other transcription factors Silencers are repressors that inhibit transcription Control sequences Promoter Enhancer Related genes located on different chromosomes can be controlled by similar enhancer sequences Copyright 2009 Pearson Education, Inc.

28 11.5 Complex assemblies of proteins control eukaryotic transcription Animation: Initiation of Transcription Copyright 2009 Pearson Education, Inc.

29 Enhancers Promoter Gene DNA Transcription factors Activator proteins Other proteins RNA polymerase Bending of DNA Transcription

30 11.6 Eukaryotic RNA may be spliced in more than one way Alternative RNA splicing Production of different mrnas from the same transcript Results in production of more than one polypeptide from the same gene Can involve removal of an exon with the introns on either side Animation: RNA Processing Copyright 2009 Pearson Education, Inc.

31 Exons DNA RNA transcript RNA splicing or mrna

32 11.7 Small RNAs play multiple roles in controlling gene expression RNA interference (RNAi) Prevents expression of a gene by interfering with translation of its RNA product Involves binding of small, complementary RNAs to mrna molecules Leads to degradation of mrna or inhibition of translation MicroRNA Single-stranded chain about 20 nucleotides long Binds to protein complex MicroRNA + protein complex binds to complementary mrna to interfere with protein production Copyright 2009 Pearson Education, Inc.

33 11.7 Small RNAs play multiple roles in controlling gene expression Animation: Blocking Translation Animation: mrna Degradation Copyright 2009 Pearson Education, Inc.

34 Protein mirna 1 2 Target mrna mirnaprotein complex 3 4 mrna degraded OR Translation blocked

35 11.8 Translation and later stages of gene expression are also subject to regulation Control of gene expression also occurs with Breakdown of mrna Initiation of translation Protein activation Protein breakdown Copyright 2009 Pearson Education, Inc.

36 Folding of polypeptide and formation of S S linkages Cleavage Initial polypeptide (inactive) Folded polypeptide (inactive) Active form of insulin

37 11.9 Review: Multiple mechanisms regulate gene expression in eukaryotes Many possible control points exist; a given gene may be subject to only a few of these Chromosome changes (1) DNA unpacking Control of transcription (2) Regulatory proteins and control sequences Control of RNA processing Addition of 5 cap and 3 poly-a tail (3) Splicing (4) Flow through nuclear envelope (5) Copyright 2009 Pearson Education, Inc.

38 11.9 Review: Multiple mechanisms regulate gene expression in eukaryotes Many possible control points exist; a given gene may be subject to only a few of these Breakdown of mrna (6) Control of translation (7) Control after translation Cleavage/modification/activation of proteins (8) Breakdown of protein (9) Copyright 2009 Pearson Education, Inc.

39 11.9 Review: Multiple mechanisms regulate gene expression in eukaryotes Applying Your Knowledge For each of the following, determine whether an increase or decrease in the amount of gene product is expected The mrna fails to receive a poly-a tail during processing in the nucleus The mrna becomes more stable and lasts twice as long in the cell cytoplasm The region of the chromatin containing the gene becomes tightly compacted An enzyme required to cleave and activate the protein product is missing Copyright 2009 Pearson Education, Inc.

40 11.9 Review: Multiple mechanisms regulate gene expression in eukaryotes Animation: Protein Degradation Animation: Protein Processing Copyright 2009 Pearson Education, Inc.

41 NUCLEUS Chromosome DNA unpacking Other changes to DNA Gene RNA transcript Transcription Exon Intron Addition of cap and tail Gene mrna in nucleus Splicing Cap Tail Flow through nuclear envelope mrna in cytoplasm CYTOPLASM Breakdown of mrna Translation Brokendown mrna Polypeptide Cleavage / modification / activation Active protein Breakdown of protein Brokendown protein

42 NUCLEUS Chromosome DNA unpacking Other changes to DNA Gene RNA transcript Transcription Exon Intron Addition of cap and tail Gene Splicing Tail mrna in nucleus Cap Flow through nuclear envelope

43 mrna in cytoplasm CYTOPLASM Breakdown of mrna Translation Brokendown mrna Polypeptide Cleavage / modification / activation Active protein Breakdown of protein Brokendown protein

44 11.10 Cascades of gene expression direct the development of an animal Role of gene expression in fruit fly development Orientation from head to tail Maternal mrnas present in the egg are translated and influence formation of head to tail axis Segmentation of the body Protein products from one set of genes activate other sets of genes to divide the body into segments Production of adult features Homeotic genes are master control genes that determine the anatomy of the body, specifying structures that will develop in each segment Copyright 2009 Pearson Education, Inc.

45 11.10 Cascades of gene expression direct the development of an animal Animation: Cell Signaling Animation: Development of Head-Tail Axis in Fruit Flies Copyright 2009 Pearson Education, Inc.

46 Eye Antenna Leg Head of a normal fruit fly Head of a developmental mutant

47 Eye Antenna Head of a normal fruit fly

48 Leg Head of a developmental mutant

49 Egg cell within ovarian follicle Follicle cells 1 Egg cell Protein signal Gene expression Head mrna Embryo 3 2 Cascades of gene expression Body segments Adult fly Gene expression 4

50 11.11 CONNECTION: DNA microarrays test for the transcription of many genes at once DNA microarray Contains DNA sequences arranged on a grid Used to test for transcription mrna from a specific cell type is isolated Fluorescent cdna is produced from the mrna cdna is applied to the microarray Unbound cdna is washed off Complementary cdna is detected by fluorescence Copyright 2009 Pearson Education, Inc.

51 DNA microarray Each well contains DNA from a particular gene Actual size (6,400 genes) 1 mrna isolated Reverse transcriptase and fluorescent DNA nucleotides 2 cdna made from mrna 3 4 Unbound cdna rinsed away cdna applied to wells cdna Fluorescent spot Nonfluorescent spot DNA of an expressed gene DNA of an unexpressed gene

52 11.12 Signal transduction pathways convert messages received at the cell surface to responses within the cell Signal transduction pathway is a series of molecular changes that converts a signal at the cell s surface to a response within the cell Signal molecule is released by a signaling cell Signal molecule binds to a receptor on the surface of a target cell Copyright 2009 Pearson Education, Inc.

53 11.12 Signal transduction pathways convert messages received at the cell surface to responses within the cell Relay proteins are activated in a series of reactions A transcription factor is activated and enters the nucleus Specific genes are transcribed to initiate a cellular response Animation: Overview of Cell Signaling Animation: Signal Transduction Pathways Copyright 2009 Pearson Education, Inc.

54 Signaling cell 1 2 Signaling molecule Receptor protein 3 Plasma membrane Target cell Relay proteins Transcription factor (activated) 4 Nucleus DNA 5 mrna Transcription New protein 6 Translation

55 11.13 EVOLUTION CONNECTION: Cellsignaling systems appeared early in the evolution of life Yeast mating is controlled by a signal transduction pathway Yeast have two mating types: a and Each produces a chemical factor that binds to receptors on cells of the opposite mating type Binding to receptors triggers growth toward the other cell and fusion Cell signaling processes in multicellular organisms are adaptations of those in unicellular organisms such as bacteria and yeast Copyright 2009 Pearson Education, Inc.

56 Receptor a factor Yeast cell, mating type a a factor Yeast cell, mating type a a/

57 CLONING OF PLANTS AND ANIMALS Copyright 2009 Pearson Education, Inc.

58 11.14 Plant cloning shows that differentiated cells may retain all of their genetic potential Most differentiated cells retain a full set of genes, even though only a subset may be expressed Evidence is available from Plant cloning A root cell can divide to form an adult plant Animal limb regeneration Remaining cells divide to form replacement structures Involved dedifferentiation followed by redifferentiation into specialized cells Copyright 2009 Pearson Education, Inc.

59 Root of carrot plant Single cell Root cells cultured in nutrient medium Cell division in culture Plantlet Adult plant

60 11.15 Nuclear transplantation can be used to clone animals Nuclear transplantation Replacing the nucleus of an egg cell or zygote with a nucleus from an adult somatic cell Early embryo (blastocyst) can be used in Reproductive cloning Implant embryo in surrogate mother for development New animal is genetically identical to nuclear donor Therapeutic cloning Remove embryonic stem cells and grow in culture for medical treatments Induce stem cells to differentiate Copyright 2009 Pearson Education, Inc.

61 Donor cell Nucleus from donor cell Reproductive cloning Implant blastocyst in surrogate mother Clone of donor is born Remove nucleus from egg cell Add somatic cell from adult donor Grow in culture to produce an Therapeutic early embryo cloning (blastocyst) Remove embryonic stem cells from blastocyst and grow in culture Induce stem cells to form specialized cells

62 Donor cell Nucleus from donor cell Remove nucleus from egg cell Add somatic cell from adult donor Grow in culture to produce an early embryo (blastocyst)

63 Reproductive cloning Implant blastocyst in surrogate mother Clone of donor is born Therapeutic cloning Remove embryonic stem cells from blastocyst and grow in culture Induce stem cells to form specialized cells

64 11.16 CONNECTION: Reproductive cloning has valuable applications, but human reproductive cloning raises ethical issues Cloned animals can show differences from their parent due to a variety of influences during development Reproductive cloning is used to produce animals with desirable traits Agricultural products Therapeutic agents Restoring endangered animals Human reproductive cloning raises ethical concerns Copyright 2009 Pearson Education, Inc.

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66 11.17 CONNECTION: Therapeutic cloning can produce stem cells with great medical potential Stem cells can be induced to give rise to differentiated cells Embryonic stem cells can differentiate into a variety of types Adult stem cells can give rise to many but not all types of cells Therapeutic cloning can supply cells to treat human diseases Research continues into ways to use and produce stem cells Copyright 2009 Pearson Education, Inc.

67 Blood cells Adult stem cells in bone marrow Cultured embryonic stem cells Different culture conditions Nerve cells Heart muscle cells Different types of differentiated cells

68 THE GENETIC BASIS OF CANCER Copyright 2009 Pearson Education, Inc.

69 11.18 Cancer results from mutations in genes that control cell division Mutations in two types of genes can cause cancer Oncogenes Proto-oncogenes normally promote cell division Mutations to oncogenes enhance activity Tumor-suppressor genes Normally inhibit cell division Mutations inactivate the genes and allow uncontrolled division to occur Copyright 2009 Pearson Education, Inc.

70 11.18 Cancer results from mutations in genes that control cell division Oncogenes Promote cancer when present in a single copy Can be viral genes inserted into host chromosomes Can be mutated versions of proto-oncogenes, normal genes that promote cell division and differentiation Converting a proto-oncogene to an oncogene can occur by Copyright 2009 Pearson Education, Inc. Mutation causing increased protein activity Increased number of gene copies causing more protein to be produced Change in location putting the gene under control of new promoter for increased transcription

71 11.18 Cancer results from mutations in genes that control cell division Tumor-suppressor genes Promote cancer when both copies are mutated Copyright 2009 Pearson Education, Inc.

72 Proto-oncogene DNA Mutation within the gene Multiple copies of the gene Gene moved to new DNA locus, under new controls Oncogene New promoter Hyperactive growthstimulating protein in normal amount Normal growthstimulating protein in excess Normal growthstimulating protein in excess

73 Tumor-suppressor gene Mutated tumor-suppressor gene Normal growthinhibiting protein Defective, nonfunctioning protein Cell division under control Cell division not under control

74 11.19 Multiple genetic changes underlie the development of cancer Four or more somatic mutations are usually required to produce a cancer cell One possible scenario for colorectal cancer includes Activation of an oncogene increases cell division Inactivation of tumor suppressor gene causes formation of a benign tumor Additional mutations lead to a malignant tumor Copyright 2009 Pearson Education, Inc.

75 Colon wall Cellular changes: 1 Increased cell division 2 3 Growth of polyp Growth of malignant tumor (carcinoma) DNA changes: Oncogene activated Tumor-suppressor gene inactivated Second tumorsuppressor gene inactivated

76 Chromosomes 1 mutation 2 mutations 3 mutations 4 mutations Normal cell Malignant cell

77 11.20 Faulty proteins can interfere with normal signal transduction pathways Path producing a product that stimulates cell division Product of ras proto-oncogene relays a signal when growth hormone binds to receptor Product of ras oncogene relays the signal in the absence of hormone binding, leading to uncontrolled growth Copyright 2009 Pearson Education, Inc.

78 11.20 Faulty proteins can interfere with normal signal transduction pathways Path producing a product that inhibits cell division Product of p53 tumor-suppressor gene is a transcription factor p53 transcription factor normally activates genes for factors that stop cell division In the absence of functional p53, cell division continues because the inhibitory protein is not produced Copyright 2009 Pearson Education, Inc.

79 Growth factor Target cell Normal product of ras gene Relay proteins Receptor Hyperactive relay protein (product of ras oncogene) issues signals on its own Transcription factor (activated) DNA Nucleus Transcription Protein that Stimulates cell division Translation

80 Growth-inhibiting factor Receptor Relay proteins Transcription factor (activated) Nonfunctional transcription factor (product of faulty p53 tumor-suppressor gene) cannot trigger transcription Normal product of p53 gene Transcription Protein that inhibits cell division Translation Protein absent (cell division not inhibited)

81 11.21 CONNECTION: Lifestyle choices can reduce the risk of cancer Carcinogens are cancer-causing agents that damage DNA and promote cell division X-rays and ultraviolet radiation Tobacco Healthy lifestyle choices Avoiding carcinogens Avoiding fat and including foods with fiber and antioxidants Regular medical checkups Copyright 2009 Pearson Education, Inc.

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83 A typical operon DNA Regulatory gene Promoter Operator Gene 1 Gene 2 Gene 3 Encodes repressor that in active form attaches to operator RNA polymerase binding site Switches operon on or off

84 Nucleus from donor cell Early embryo resulting from nuclear transplantation Surrogate mother Clone of donor

85 Nucleus from donor cell Early embryo resulting from nuclear transplantation Embryonic stem cells in culture Specialized cells

86 controlled by protein called operons prokaryotic genes often grouped into Gene regulation (a) is a normal gene that can be mutated to an in eukaryotes may involve when oncogene abnormal may lead to can cause (b) in active form binds to are switched on/off by (c) (d) (e) (f) (g) occurs in female mammals are proteins that promote transcription can produce multiple mrnas per gene

87 You should now be able to 1. Explain how prokaryotic gene control occurs in the operon 2. Describe the control points in expression of a eukaryotic gene 3. Describe DNA packing and explain how it is related to gene expression 4. Explain how alternative RNA splicing and micrornas affect gene expression 5. Compare and contrast the control mechanisms for prokaryotic and eukaryotic genes Copyright 2009 Pearson Education, Inc.

88 You should now be able to 6. Distinguish between terms in the following groups: promoter operator; oncogene tumor suppressor gene; reproductive cloning therapeutic cloning 7. Define the following terms: Barr body, carcinogen, DNA microarray, homeotic gene; stem cell; X-chromosome inactivation 8. Describe the process of signal transduction, explain how it relates to yeast mating, and explain how it is disrupted in cancer development Copyright 2009 Pearson Education, Inc.

89 You should now be able to 9. Explain how cascades of gene expression affect development 10. Compare and contrast techniques of plant and animal cloning 11. Describe the types of mutations that can lead to cancer 12. Identify lifestyle choices that can reduce cancer risk Copyright 2009 Pearson Education, Inc.

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