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 Overview of transcription

5 E. coli: Initiation

6 E. coli: The promoter

7 Eukaryotes: Initiation

8 Topics Gene regulation in bacteria Genetic analysis of gene induction Basic logical and molecular principles of genetic switches Gene regulation in eukaryotes Chromatin

9 Why?

10 Gene regulation in bacteria

11 Genetic Switches An/Ausschalter

12 Positive vs negative regulation

13 Modular functionality: e.g., allosteric effectors

14 Model: the lac system of E. coli Induction of ß-galactosidase by the disaccharide lactose Francois Jacob André Lwoff Jacques Monod

15 The inducer: lactose

16 Simplified lac operon model

17 No lactose present

18 Lactose present

19 Genetic dissection of lac system Genetic components I I -, I s Y Y - P P - Z Z - O O c A A -

20 Generating partial diploids in E. coli Bacterial genetics is merozygote genetics

21 O c mutants: Synthesis of ß-galactosidase/permease O c is a constitutive mutation Operator (O) is cis-acting

22 Interpretation Operators are cis-acting

23 I - mutants: Synthesis of ß- galactosidase/permease Repressor (I) is trans-acting

24 Interpretation Repressors are trans-acting

25 I s alleles: Synthesis of ß- galactosidase/permease Repressor itself is always repressed

26 Interpretation Repressor contains a lactose-binding site

27 Lactose present

28 Genetic analysis: I vs O both elements act in repressing lac operon they fundamentally differ in their mode of action (cis vs trans mode) reveals important aspects of the molecular mechanism of lac repression

29 Control of lac system: Lactose vs glucose

30 Catabolite repression of the lac operon Glucose is a catabolite of lactose camp/cap complex is an activator of transcription

31 Control of the lac operon

32 Molecular anatomy of the genetic switch

33 The operator is a specific DNA sequence Very specific sequence! One base change enough to eliminate O function

34 Many DNA binding sites are symmetrical

35 Binding of CAP bends DNA Recognition of specific CAP-binding site

36 CAP/RNA pol: Binding sites

37 Helix-turn-helix is a common DNA-binding motif Dimers Specific contacts with bases in major groove

38 AA side chains determine specificity of DNA binding Homeodomain

39 Repression vs activation

40 Genetic switches are often part of a cascade mechanism A TF B C

41 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 gene are often clustered into operons on the chro and transcribed together into 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

42 Gene regulation in eukaryotes

43 Drosophila: MSL complex and dosage compensation

44 Overview of transcriptional regulation

45 Eukaryotic promoter

46 The yeast GAL system

47 The transcriptional activator Gal4 TF: sequence-specific binding to regions outside the promoters of target genes

48 TFs: Modular Proteins

49 Transcriptional activators and the transcription machinery

50 Enhancer action: Mechanism Cooperativity Synergism

51 Disperse distribution of enhancer elements DPP of Drosophila kb

52 Modular and combinatorial control eve TATA lacz

53 Chromatin

54 Chromatin dynamics

55 Chromatin remodeling e.g., by SWI-SNF complex

56 Histone modifications and the histone code

57 Tup1, a histone deacetylase from yeast, is a corepressor

58 Linking TFs and chromatin dynamics

59 Enhanceosome Cooperativity Synergism

60 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)

61 THE END

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