Einführung in die Genetik

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1 Einführung in die Genetik Prof. Dr. Kay Schneitz (EBio Pflanzen) Prof. Dr. Claus Schwechheimer (PlaSysBiol)

2 Einführung in die Genetik - Inhalte 1 Einführung KS 2 Struktur von Genen und Chromosomen KS 3 Genfunktion KS 4 Transmission der DNA während der Zellteilung KS 5 Vererbung von Einzelgenveränderungen KS 6 Genetische Rekombination (Eukaryonten) KS 7 Genetische Rekombination (Bakterien/Viren) KS 8 Rekombinante DNA-Technologie CS 9 Kartierung/Charakterisierung ganzer Genome CS 10 Genmutationen: Ursache und Reparatur CS 11 Veränderungen der Chromosomen CS 12 Genetische Analyse biologischer Prozesse CS 13 Transposons bei Eukaryonten CS 14 Regulation der Genexpression KS 15 Regulation der Zellzahl - Onkogene CS

3 Regulation of Gene Expression Genetics 14

4 Summary Cells respond to intrinsic and extrinsic signals by modulating transcriptional control of certain genes Gene activity is the result of the function of cis- and trans-acting factors Trans-acting proteins react to environmental signals by using built-in sensors that continually monitor cellular conditions Coordinated gene regulation in bacteria genes are often clustered into operons on the chromosome and transcribed together as multigenic mrnas one cluster of regulatory sites per operon is sufficient to regulate expression of several genes Negative vs positive regulation repressor proteins bind to DNA at operator site thereby blocking transcription (e.g., lac operon) activator proteins activate transcription by binding to DNA at the promoter region (e.g., camp/cap regulation of lac operon) Molecular anatomy of genetic switch regulatory proteins have DNA-binding domains (e.g., HLH) and protein-protein interaction domains (modular specificity of gene regulation depends on specific protein-dna interactions mediated by the chemical interactions between aa side chains and chemical groups of DNA bases

5 Summary Eukaryotic gene regulation resembles bacterial gene regulation trans-acting factors binding to cis-regulatory elements on the DNA this regulatory factors determine the level of transcription by regulating the binding of RNA pol II to the promoter of a gene Enhancers/UAS cis-regulatory elements, possibly located quite far away (>10-50kb) from promoter combinatorial interactions among different transcription factors enhanceosome: complexes of regulatory proteins that interact in cooperative and synergistic fashion --> high levels of transcription through recruitment of RNA pol II Gene regulation and chromatin eukaryotic genes are packed in chromatin activation/repression requires specific modifications to chromatin genes are mostly turned off and kept silent in part by nucleosomes and condensed chromatin histone code: pattern of posttranslational modifications of histone tails (acetylation, methylation, phosphorylation etc). histone code is an epigenetic mark involved in nucleosome positioning and chromatin condensation that can be altered by TFs TFs recruit for example ATP-dependent chromatin remodelers (e.g., SWI-SNF)

6 Control of cell number - oncogenes Genetics 15

7 Tumors The cell cycle The ubiquitin-proteasome system Apoptosis Cancerogenesis

8 Tumors uncontrolled cell divisions and growth invasion and colonization of tissue malignant vs. benign tumors metastasis

9 Tumor types carcinoma - epithelial cell cancer sarcoma - connective tissue or muscle cancer leukemia or lymphoma - blood cell cancers others

10 Tumors - incidence rates

11 Tumors - types and distribution

12 The cell cycle

13 Cell cycle studies FACS sorting (fluorescence activated cell sorting)

14 Cell cycle studies Northern blot Western blot

15 Cell cycle mutants of yeast

16 Temperature sensitive mutants in cell cycle analysis

17 Major check points in cell cycle control

18 Cell cycle studies Northern blot Western blot

19 Cyclins and cyclin-dependent kinases are differentially transcribed throughout the cell cycle

20 Cyclins and cyclin-dependent kinases (CDKs) Cyclin-CDK! Vertebrates!! Yeast! Complex!Cyclin! Cdk!!Cyclin! Cdk! G1-Cdk!!Cyclin D Cdk4, Cdk6!Cln3! Cdk1! G1/S-Cdk!Cyclin E! Cdk2!!Cln1,2! Cdk1! S-Cdk!!Cyclin A! Cdk2!!Clb5,6! Cdk1! M-Cdk!!Cyclin B! Cdk1!!Clb1,2,3,4 Cdk1!

21 Cyclin-CDK complexes control the cell cycle

22 Mitosis promoting factor and cyclins

23 Cell cycle control by Cyclin-CDKs

24 Cell cycle control by Cyclin-CDKs

25 Cyclins and CDKs in cell cycle control

26 Mitotic cyclins and protein phosphorylation

27 The ubiquitin-proteasome system and protein degradation

28 The ubiquitination machinery E1 ubiquitin activating emzyme E2 ubiquitin conjugating emzyme E3 ubiquitin ligase

29 The ubiquitination machinery

30 The 26S proteasome

31 Mitotic cyclins and protein degradation

32 The anaphase promoting complex (APC)

33 Cancerogenesis

34 Major check points in cell cycle control

35 Examples for receptors in signaling

36 Receptor tyrosine kinases JAK/STAT pathway

37 Receptor tyrosine kinases Growth factors (EGF, TGF etc.)

38 Ras

39 Ras

40 The Retinoblastoma (RB) tumor suppressor

41 Retinoblastoma protein (Rb) blocks E2F

42 Prelavance of p53 tumour-suppressor mutations

43 p53 is phosphorylated in response to DNA damage

44 p53 phosphorylation blocks the cell cycle

45 p53 phosphorylation blocks the cell cycle p53 +/+ days

46 Mutations can induce cancer Ras Rb p53

47 and invasive cell divisions and cell growth Summary Tumours are the result of uncontrolled The cell cycle is governed by cyclins and cyclin-dependent kinases Cyclins and cyclin-dependent kinases are under transcriptional control Cyclins are degraded by the UPS E1, E2, E2 enzymes and ubiquitin 26S proteasome Apoptosis regulated cell number Apoptosis is a controlled process p53, RB and Ras are important cell cycle regulators

48 The end

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