1. Unit 4 Proteins Transcription (DNA to mRNA) Translation (mRNA to tRNA to proteins) Gene expression/regulation (turning genes on and off) Viruses 1

2. Yesterday’s Exit Ticket Template DNA3’3’TACAATGCATTG__’ Non-Template 5’5’ATGTTACGTAAC__’ mRNA5’AUGUUACGUAAC3’ 1)Fill in the blanks. 2)What is the amino acid sequence corresponding to the DNA and RNA sequences below? 2 Met-Leu-Arg-Asn

3. Today’s Plan What is gene regulation? Why do cells do it? How genes are regulated – Bacteria – Eukaryotes Mechanisms of Development 3

4. Is the DNA in this cell Different From this one? Or this one? 4

5. GENE REGULATION What is gene regulation? Turning genes on and off in the right time and place Controlling the quantity of proteins produced 5

6. GENE REGULATION Why do genes need to be regulated? Every cell in our body has the same set of genes Our body consists of trillions of cells and millions of distinct cell types.  What makes a skin cell different from a liver cell? 6

7. Why do genes need to be regulated? 1.Differentiated structures/organs 2.Not all of a single cell’s functions are needed all the time (e.g. we shouldn’t make insulin constantly) 7

8. Why do genes need to be regulated? 1.Differentiated structures/organs 2.Not all of a single cell’s functions are needed all the time (e.g. we shouldn’t make insulin constantly) 3.Waste of energy/molecules to express genes whose products are not needed 8

9. Why do genes need to be regulated? 1.Differentiated structures/organs 2.Not all of a single cell’s functions are needed all the time (e.g. we shouldn’t make insulin constantly) 3.Waste of energy/molecules to express genes whose products are not needed 4.Some functions are mutually exclusive; two enzymes may have opposite functions 9

10. Examples: different cell types within a human Muscle cells express muscle actin and myosin Hair and nail cells express keratin Blood cells express hemoglobin 10

11. Today’s Plan What is gene regulation? Why do cells do it? How genes are regulated – Bacteria – Eukaryotes Mechanisms of Development 11

12. “…Consider, for instance, an individual E. coli cell living in the...human colon, dependent for its nutrients on the whimsical eating habits of its host” textbook, p. 352 tryptophan present? YE S no need to synthesiz e tryptoph an NO need to synthesize tryptophan express genes for tryptophan synthesis HOW? 12

13. How can a cell do this, with no “brain” or “intelligence” directing it?  Molecules can act as signals by directly influencing transcription 13

14. How does this really work? Example: E. coli regulation of tryptophan (1) turning multiple genes “on” and “off” (2) doing so at the appropriate times 14

15. Sweatblock.com; netropolus.com Negative Feedback If something is present, don’t make more of it! - 15

16. Gene regulation in bacteria Operator: the “on-off switch” that controls the access of RNA polymerase to the genes Operon: promoter, operator, and genes 16

17. (2) expressing the genes of the operon at the right time For the trp operon, RNA polymerase can bind when nothing is bound to the operator 17

18. (2) expressing the genes of the operon at the right time 18

19. (2) expressing the genes of the operon at the right time A system of negative feedback 19

20. (2) expressing the genes of the operon at the right time How do we stop repression? Binding of tryptophan to repressor is reversible Repressor activation/deactivation depends upon relative concentrations of tryptophan and repressor protein Binding of tryptophan to repressor is reversible Repressor activation/deactivation depends upon relative concentrations of tryptophan and repressor protein 20

21. Today’s Plan What is gene regulation? Why do cells do it? How genes are regulated – Bacteria – Eukaryotes Mechanisms of Development 21

22. Eukaryotes: Differential Gene Expression Differences between cell types result from differential gene expression (a) Fertilized eggs of a frog (b) Newly hatched tadpole 22

23. Fig. 18-6 DNA Signal Gene NUCLEUS Chromatin modification Chromatin Gene available for transcription Exon Intron Tail RNA Cap RNA processing Primary transcript mRNA in nucleus Transport to cytoplasm mRNA in cytoplasm Translation CYTOPLASM Degradation of mRNA Protein processing Polypeptide Active protein Cellular function Transport to cellular destination Degradation of protein Transcription In eukaryotes, gene expression can be regulated at many different stages. 23

24. Fig. 18-6a DNA Signal Gene NUCLEUS Chromatin modification Chromatin Gene available for transcription Exon Intron Tail RNA Cap RNA processing Primary transcript mRNA in nucleus Transport to cytoplasm CYTOPLASM Transcription 24

25. Fig. 18-6b mRNA in cytoplasm Translation CYTOPLASM Degradation of mRNA Protein processing Polypeptide Active protein Cellular function Transport to cellular destination Degradation of protein 25

26. How is transcription regulated in eukaryotes? Promoters and introns aren’t the only non-coding regions of DNA! Meet the Enhancers: http://bja.oxfordjournals.org 26

27. How is transcription regulated in eukaryotes? Liver enhancerLiver gene (enzyme) Lens gene (crystallin)Lens enhancer Promoter Meet the Enhancers: DNA control regions corresponding to a specific gene Can be upstream, downstream, or in an intron Comprised of control elements Control Elements 27

28. How is transcription regulated in eukaryotes? Liver enhancerLiver gene (enzyme) Lens gene (crystallin)Lens enhancer Promoter How does a DNA region control a gene? Through activators specific to each control element Activators are specialized transcription factors (proteins) Control Elements 28

29. Would you go down this: sfomom.blogspot.com; sabotagetimes.com Without this? 29

30. Stopped Editing Here 30

31. Liver enhancerLiver gene (enzyme) Lens gene (crystallin)Lens enhancer Liver cellLens cell Lens activatorsLiver activators Pro. Lens gene ON Liver gene ON Lens gene OFF Liver gene OFF Animation 31

32. Today’s Plan What is gene regulation? Why do cells do it? How genes are regulated – Bacteria – Eukaryotes Mechanisms of Development – Example of malformed frogs 32

33. Development 1.Determination and Differentiation 2.Getting the right parts in the right places: Pattern formation 33

34. embryonic development Zygote 34

35. (a) Fertilized eggs of a frog (b) Newly hatched tadpole one zygote    many cell types, tissues, organs cell division (mitosis), differentiation, morphogenesis The key is differential gene regulation 1. Differentiation and determination 35

36. The process of development: some useful terms 1. Differentiation and determination When I grow up I will be a heart cell! Better start making actin and myosin! Hello, my name is Celly Hello, my name is Celly You’re a heart cell too! Yay! Hello, my name is Celly Hello, my name is Cella 36

37. 37

38. TWO COMPLEMENTARY MECHANISMS of DIFFERENTIATION Cytoplasmic Determinants  before fertilization, when eggs are made  maternally derived Inductive Signals  once there are multiple cells  substance from outside a cell (e.g., signal from nearby cell) influences cell’s gene expression 1. Differentiation and determination 38

39. (a) Cytoplasmic determinants Two different cytoplasmic determinants Unfertilized egg Sperm Fertilization Zygote Mitotic cell division Two-celled embryo Nucleus 39

40. (b) Induction by nearby cells Signal molecule (inducer) Chain reaction Early embryo NUCLEUS Signal receptor 1. Differentiation and determination 40

41. Embryonic precursor cell Nucleus OFF DNA master regulatory gene myoD Other muscle-specific genes OFF has potential to develop into a variety of different cell types 41

42. OFF mRNA MyoD protein (transcription factor) Muscle cell precursor (determined) Embryonic precursor cell Nucleus OFF DNA Other muscle-specific genes OFF master regulatory gene myoD 42

43. mRNA muscle proteins Part of a muscle fiber (fully differentiated cell) MyoD Another transcription factor OFF mRNA MyoD protein (transcription factor) Embryonic precursor cell Nucleus OFF DNA Other muscle-specific genes OFF Muscle cell precursor (determined) master regulatory gene myoD 43

44. Outline 1.Determination and Differentiation 2.Getting the right parts in the right places: Pattern formation 44

45. Antenna Mutant Wild type Eye Leg How do you get the right tissues/organs in the right places??? 45

46. Pattern Formation: Setting Up the Body Plan Pattern formation = development of spatial organization of tissues and organs  establishment of major body axes Positional information = molecular cues that tell a cell its location 46

47. Cytoplasmic determinants in eggs are products of maternal effect genes (a.k.a. egg polarity genes) Unfertilized egg unevenly distributed RNA, proteins (from mother) 47

48. A closer look at the formation of the anterior-posterior (head-tail) axis: the bicoid gene Presence of bicoid protein = “put head here” OR Lack of bicoid protein = “head does not go here” 48

49. Cytoplasmic determinants in eggs are products of maternal effect genes (a.k.a. egg polarity genes) Unfertilized egg unevenly distributed RNA, proteins (from mother) mutation in maternal effect gene 49

50.  Gradient of bicoid protein determines anterior- posterior axis 50

51. Today’s Take-Homes: Gene expression can be regulated at many stages Transcription regulation is a common mechanism Eukaryotes use enhancers and activators for differential gene expression Protein concentrations from mom’s egg play a key role in development 51

52. Today’s Exit Ticket Describe, in your own words, the role of enhancers and activators in eukaryotic gene regulation. 52