1. AP Biology Discussion Notes 2/25/2015

2. Goals for Today Be able to describe regions of DNA and how they are important to gene expression in Bacteria (Prokaryotes) & Eukaryotes

3. Question of the Day 2/25 What two things would be found on a DNA sequence before (______ stream of) a coding region or genes?

4. Conducting the Genetic Orchestra CH18 Prokaryotes and eukaryotes alter gene expression in response to their changing environment

5. 18.1: Bacteria often respond to environmental change by regulating transcription Natural selection has favored bacteria that produce only the products needed by that cell A cell can regulate the production of enzymes by feedback inhibition or by gene regulation Gene expression in bacteria is controlled by the operon model

6. Repressible and Inducible Operons: Two Types of Negative Gene Regulation An inducible operon is one that is usually ____; a molecule called an inducer inactivates the repressor and turns on transcription What is the “inducer” in the Lac Operon?

7. Figure 18.4a (a) Lactose absent, repressor active, operon off Regulatory gene Promoter Operator DNA lacZ lac I DNA mRNA 5 3 No RNA made RNA polymerase Active repressor Protein

8. Figure 18.4b (b) Lactose present, repressor inactive, operon on lac I lac operon lacZlacYlacADNA mRNA 5 3 Protein mRNA 5 Inactive repressor RNA polymerase Allolactose (inducer)  -Galactosidase PermeaseTransacetylase

9. Positive Gene Regulation Some operons are also subject to positive control through a stimulatory protein, such as catabolite activator protein (CAP), an activator of transcription When glucose (a preferred food source of E. coli) is scarce, CAP is activated by binding with cyclic AMP (cAMP) Activated CAP attaches to the promoter of the lac operon and increases the affinity of RNA polymerase, thus accelerating transcription

10. When glucose levels increase, CAP detaches from the lac operon, and transcription returns to a normal rate CAP helps regulate other operons that encode enzymes used in catabolic pathways Positive Gene Regulation

11. Figure 18.5a Promoter DNA CAP-binding site lacZ lac I RNA polymerase binds and transcribes Operator cAMP Active CAP Inactive CAP Allolactose Inactive lac repressor (a) Lactose present, glucose scarce (cAMP level high): abundant lac mRNA synthesized

12. Figure 18.5b Promoter DNA CAP-binding site lacZ lac I Operator RNA polymerase less likely to bind Inactive lac repressor Inactive CAP (b) Lactose present, glucose present (cAMP level low): little lac mRNA synthesized

13. Fill in the blanks

14. Repressible and Inducible Operons: Two Types of Negative Gene Regulation A repressible operon is one that is usually ____; binding of a repressor to the operator shuts off transcription The trp operon is a repressible operon

15. E. coli can synthesize the amino acid tryptophan Think of these questions and think about energy and natural selection: –When would “want” to do this? –When would “not want” to do this? trp OPERON

16. By default the trp operon is on; and the genes for tryptophan synthesis are transcribed When tryptophan is present, it binds to the trp repressor protein, which turns the operon off trp OPERON

17. The repressor can be in an active or inactive form, depending on the presence of other molecules The repressor is active only in the presence of its corepressor tryptophan; –A corepressor is a molecule that cooperates with a repressor protein to switch an operon off thus the trp operon is turned off (repressed) if tryptophan levels are high trp OPERON

18. Figure 18.3a Promoter DNA Regulatory gene mRNA trpR 5 3 Protein Inactive repressor RNA polymerase Promoter trp operon Genes of operon Operator mRNA 5 Start codonStop codon trpEtrpDtrpC trpB trpA EDCBA Polypeptide subunits that make up enzymes for tryptophan synthesis (a) Tryptophan absent, repressor inactive, operon on

19. Figure 18.3b-1 (b) Tryptophan present, repressor active, operon off DNA mRNA Protein Tryptophan (corepressor) Active repressor

20. Figure 18.3b-2 (b) Tryptophan present, repressor active, operon off DNA mRNA Protein Tryptophan (corepressor) Active repressor No RNA made

21. Precursor Feedback inhibition Enzyme 1 Enzyme 2 Enzyme 3 Tryptophan (a) (b) Regulation of enzyme activity Regulation of enzyme production Regulation of gene expression   trpE gene trpD gene trpC gene trpB gene trpA gene Figure 18.2

22. Promoter DNA Regulatory gene mRNA trpR 5 3 Protein Inactive repressor RNA polymerase Promoter trp operon Genes of operon Operator mRNA 5 Start codonStop codon trpEtrpDtrpC trpB trpA EDCBA Polypeptide subunits that make up enzymes for tryptophan synthesis (a) Tryptophan absent, repressor inactive, operon on (b) Tryptophan present, repressor active, operon off DNA mRNA Protein Tryptophan (corepressor) Active repressor No RNA made Figure 18.3

23. Repressible and Inducible Operons: Two Types of Negative Gene Regulation A repressible operon is one that is usually ____ An inducible operon is one that is usually ____ The trp operon is a(n) ____________ operon The Lac operon is a(n) ____________ operon

24. Conducting the Genetic Orchestra CH18 Prokaryotes and eukaryotes alter gene expression in response to their changing environment In multicellular eukaryotes, gene expression regulates development and is responsible for differences in cell types

25. Figure 18.1

26. Eukaryotic gene expression: regulated at many stages All organisms must regulate which genes are expressed at any given time In multicellular organisms regulation of gene expression is essential for cell specialization

27. Figure 11.22 Interdigital tissue Cells undergoing apoptosis Space between digits 1 mm

28. Differential Gene Expression Almost all the cells in an organism are genetically identical Differences between cell types result from differential gene expression, – the expression of different genes by cells with the same genome Abnormalities in gene expression can lead to diseases including cancer Gene expression is regulated at many stages

29. Figure 18.6 Signal NUCLEUS Chromatin Chromatin modification: DNA unpacking involving histone acetylation and DNA demethylation DNA Gene Gene available for transcription RNA Exon Primary transcript Transcription Intron RNA processing Cap Tail mRNA in nucleus Transport to cytoplasm CYTOPLASM mRNA in cytoplasm Translation Degradation of mRNA Polypeptide Protein processing, such as cleavage and chemical modification Active protein Degradation of protein Transport to cellular destination Cellular function (such as enzymatic activity, structural support)

30. Figure 18.6a Signal NUCLEUS Chromatin Chromatin modification: DNA unpacking involving histone acetylation and DNA demethylation DNA Gene Gene available for transcription RNA Exon Primary transcript Transcription Intron RNA processing Cap Tail mRNA in nucleus Transport to cytoplasm CYTOPLASM

31. Figure 18.6b CYTOPLASM mRNA in cytoplasm Translation Degradation of mRNA Polypeptide Protein processing, such as cleavage and chemical modification Active protein Degradation of protein Transport to cellular destination Cellular function (such as enzymatic activity, structural support)

32. DNA Methylation DNA methylation, the addition of methyl groups to certain bases in DNA, is associated with reduced transcription in some species DNA methylation can cause long-term inactivation of genes in cellular differentiation

33. DNA Methylation In genomic imprinting, methylation regulates expression of either the maternal or paternal alleles of certain genes at the start of development

34. Histone Modifications In histone acetylation, acetyl groups are attached to positively charged lysines in histone tails This loosens chromatin structure, thereby promoting the initiation of transcription The addition of methyl groups (methylation) can condense chromatin; the addition of phosphate groups (phosphorylation) next to a methylated amino acid can loosen chromatin

35. Figure 18.7 Amino acids available for chemical modification Histone tails DNA double helix Nucleosome (end view) (a) Histone tails protrude outward from a nucleosome Unacetylated histones Acetylated histones (b) Acetylation of histone tails promotes loose chromatin structure that permits transcription

36. Operon Practice If you can draw it you understand it

38. Registration Materials See Ms. Benz in A156

39. Thoughts on taking the AP TEST You have worked hard all semester, and you should get your reward for all that hard work!

40. Thoughts on taking the AP TEST IF biology major: –You should take the test which will count as an elective credit and be much cheaper than other credits –You will still want to take MAJORS biology at your college/university to: Understand their specific expectations & emphasis Meet professors and make good impressions for potential lab/field jobs Meet your cohort – other students who will be going through the program with you – and form friendships & study groups

41. Thoughts on taking the AP TEST IF non-science or non-biology major: –You should take the test for easy credit while it is all fresh in your mind. It will be much more difficult and more expensive to get this same credit as a college class