KBF/MBM Molecular Biology of Mutagens, Carcinogens and Cytostatics

Lecturers: Jana Kašpárková, Viktor Brabec, Martina Špundová
Lecture: 2 hours/week
Credits: 3
Winter semester
Form of course completion: exam

The course will discuss the role of mutagens in living organisms, mechanisms underlying their reactions with nucleic acids and mechanisms of repair of the damage induced by their action. A particular chapter will be devoted to the methods used in the studies of interactions of mutagens with nucleic acids. The issues will be divided in the following way:

• Chemical compounds as mutagens and caricinogens. Definition of mutations, origin and types of mutations.
• Basic properties of nucleic acids: Chemical composition, structure and properties of nucleic acids.
• Mechanism of transfer of genetic information and its alterationsus (replication and transcription), defects in these processes and origin of mutations.
• Genetic material handling, isolation of nucleic acids and manipulation with nucleic acids.
• Determination of nucleotide sequence of nucleic acids (endonucleases, exonucleases, restriction enzymes, chemical /Maxam-Gilbert/ method, the chain-termination /Sanger/ method)  how to find out that mutation occurred.
• Reactions of mutagens, carcinogens and cytostatics: direct reactions, metabolic activation; types of DNA damage.
• Repair of damaged DNA in mammalian cells. I. total repair, excision repair: base excision repair, nucleotide excision repair, mismatch repair.
• Repair of damaged DNA in mammalian cells. II: repair of double-strand breaks, tolerant repair.
• DNA damage and cancer. Carcinogenesis.
• Current approaches in the design of new cytostatics and their development.
• Cytostatics - types of the most frequently used cytostatics, determination of the target.
• In vitro cytotoxicity tests.
• Methods and procedures for determination of mutagenicity of chemical substances and physical agents - tests on molecular, gene and chromosome level.
• Methods for studies of reactions of mutagens, carcinogens and cytostatics with DNA. I.
  a) DNA binding, detection of binding and damage,
  b) determination of sequence specificity of the binding.
• Methods for studies of reactions of mutagens, carcinogens and cytostatics with DNA. II.
  a) formation of interstrand cross-links and their quantification,
  b) alterations if conformation and stability of DNA.
• Methods for studies of reactions of mutagens, carcinogens and cytostatics with DNA. III. Reactions with cellular proteins and studies of repair processes in vitro and in cells.
• Electrochemical detection of damage of genetic material.