1. Today’s Agenda: 1. A Microarray Primer 2. Guest Speaker: Dr. Michael Schlador 3. Follow-up: the Use of Microarray Analysis in Chemotherapeutics 4. Preview of Chapter 15 & 16

2. Chapter 15: Mutations Mutations generate genetic variants These variants are then subject to recombination

3. Consequences of mutations

4. What would you predict if the mutations was….. Within a coding region Within a noncoding region

5. What are the effects of point mutations on gene products?

6. Are mutations random or induced? Luria and Delbruck used the “fluctuation test” They were interested in determining which hypothesis was correct

7. Replica plating

8. What can cause spontaneous mutations? Errors in DNA replication – Transitions: base pairs mismatch – Transversions: base pair mismatch with the normal keto base forms

9. What can cause spontaneous mutations? Errors in DNA replication – Frameshift mutations: Also known as indel mutations

10. Trinucleotide repeats may lead to disease…why?

11. How can we induce mutations? Use base analogs – Cause incorrect base pairing

12. How can we induce mutations? Use intercalating agents which mimic base pairs and slip in between the base pairs…promote indel mutations

13. How can we induce mutations? Promote base damage – UV light – Ionizing radiation Promotes large strand breaks – Aflatoxin B 1 Binds to guanine and generates an apurinic site

14. Ames Test is to determine mutagenicity

15. Biological Repair Mechanisms Direct reversal of damaged DNA Base-excision repair Nucleotide-excision repair Mismatch repair

16. Direct reversal of damaged DNA

17. Base-excision repair Carried out by DNA glycolylases, generate apurininc or apyrimidinic sites AP endonuclease nicks strand Deoxyribophosphodi- esterase removes more DNA DNA polymerase fills in the gap with new DNA

18. Nucleotide-excision repair Used to repair base damage or transcription blocks Autosomal recessive diseases, Xeroderma pigmentosum and Cockayne Syndrome result from defects in this system

19. Mismatch repair Recognize mismatch base pairs Determine which base is the incorrect one Excise the incorrect base and repair DNA

20. Error prone repair Known as translesion synthesis, requires a bypass polymerase

21. Repair of double-strand breaks What conditions can cause double strand breaks? There are two possible situations: – Nonhomologous end joining – Homologous recombination

22. Nonhomologous end joining

23. Homologous recombination Damage corrected by synthesis-dependent strand annealing (SDSA) Uses sister chromatids as templates

24. Why can mutations lead to cancer? Two types of mutations associated with cancer – Oncogenes are activated – Tumor suppressor genes are inactivated

25. Oncogenes Genes that control the cell cycle or inhibit apoptosis are considered proto- oncogenes

26. Tumor Suppressor genes Often these genes participate in regulation of cell cycle, activate cell apoptosis, or repair of damaged DNA Mutations in p53 result in 50% of all tumors

27. Chapter 16: Changes in chromosomes

28. Chromosome numbers Changes in chromosome sets are known as aberrant euploidy Changes in parts of chromosome sets is known as aneuploidy

29. Polyploids More common in plants Correlation between the number of chromosome sets and size of organism Autopolyploids: multiple chromosomes from one species Allopolyploids: sets of chromosomes from two or more different species

30. Agricultural applications Monoploid plants provide a way to select for desired traits

31. Agricultural applications Bananas are sterile triploids Autotetroploid grapes are larger than diploid grapes

32. Aneuploidy Mostly due to nondisjunction during meiosis or mitosis

33. Abnormal numbers of sex chromosome Turner Syndrome – Result of only one sex chromosome (XO)

34. Abnormal numbers of sex chromosome Klinefleter Syndrome – The result of an extra X chromosome (XXY)

35. Abnormal number of autosomes Down syndrome – Results from an extra copy of chromosome 21

36. Can Down syndrome be inherited?

37. Cancer from translocation

38. Readings for Chapter 15/16 Chapter 15: all sections except 15.5 Chapter 16: section 16.1, 16.2 Robertsonian translocation (584-585) and rearrangements and cancer (587)