1 Introduction
1.1 The Sonderforschungsbereich - an Important Instrument for Research at the Universities
1.2 Sonderforschungsbereich: A Grant System or a Concept for Basic Research
1.3 The Kernel of it all - the Projects
1.3.1 The System of E. coli as Problem and Tool
1.3.2 Membrane Lipids, Lipoproteins, and Membrane Structures
1.3.3 Resolution of Recombination Intermediates
1.3.4 Transposable Elements in Bacteria and in Plants
1.3.5 Agrobacterium-Mediated Gene Transfer to Plants
1.3.6 Resistance Mechanisms in Plants
1.3.7 Neurogenesis in Drosophila
1.3.8 Differentiation, Pattern Formation, and Sexual Induction in Volvox carteri
1.3.9 Selection of the Receptor Repertoire in the Immune System
1.3.10 Viruses as Troyan Horses in Molecular Biology
1.3.11 Alzheimer's Disease
1.4 Independent Investigators - a Novel Element in the Sonderforschungsbereich 74
1.5 The Next Grant Application is Due
1.6 The Future of the Sonderforschungsbereich 74
1.7 The Direction of Research in Molecular Biology in the Nineties
2 Molecular Mechanisms
2.1 The lac System of E. coli as Problem and Tool
Benno Muller-Hill
2.1.1 Introduction: The lac System as a Paradigm of Molecular Biology
2.1.2 The lac Repressor lac Operator System
2.1.2.1 The Primary Structure of lac and gal Repressor
2.1.2.2 The Quarternary Structure of lac Repressor
2.1.2.3 The Mechanism of Repression of the lac System
2.1.2.4 Recognition of lac Operator by lac Repressor
2.1.2.5 Replacing lac Operator by Other Targets: The Targets of trp Repressor, gal4 Protein and Putative Poly GCn Binding Protein
2.1.3 The lac Z ([beta]-galactosidase) Gene
2.1.3.1 Structure of the lac Z, the gal ET Genes of E. coli
2.1.3.2 The lac Z Gene as a Part of Cloning Vectors
2.1.4 Structure and Functional Analysis of lac Permease
2.1.5 Amyloid Precursor - Leaving the lac System for an Excursion into the Molecular Biology of Alzheimer's Disease
2.1.6 Beyond Molecular Biology: Excursions into Philosophy and History of Science
2.1.7 Summary and Outlook
2.1.8 References
2.2 Of Lipids and Membranes
Peter Overath
2.3 Studies on Regulation of Gene Expression and on Membrane Biosynthesis in Escherichia coli
Ruth Ehring
2.3.1 Summary
2.3.2 Regulation of Expression of the Galactose Operon in vitro
2.3.3 Messenger RNA-Directed Enzyme Synthesis and Changes of RNA Conformation Affecting Translational Efficiency
2.3.4 Study of Polar Effects of Insertion Mutations
2.3.5 Identification of an IS4 Encoded Protein
2.3.6 Identification of Proteins Predicted from DNA Sequences
2.3.7 E. coli Lactose Permease is Synthesized Without Cleavable Signal Sequence
2.3.8 Studies on Membrane Insertion of E. coli Lactose Permease
2.3.9 DNA/Protein Interactions in the Control of the Galactose Operon: RNA Polymerase and GAL Repressor Induce DNA-Bending
2.3.10 References
2.4 The Beginnings of Protein Engineering
Bernd Gutte
2.4.1 Summary
2.4.2 Truncated, Biologically Active Ribonuclease A
2.4.2.1 Introduction
2.4.2.2 Results
2.4.2.3 Conclusions
2.4.3 Combination of the A and B Chain of Insulin on an Anti-Insulin Antibody Column
2.4.3.1 Introduction
2.4.3.2 Results
2.4.3.3 Conclusions
2.4.4 References
2.5 Membrane Lipids, Lipoproteins and Membrane Structures. The Targets of 15 Years of Biochemical Research
Wilhelm Stoffel
2.5.1 Preface
2.5.2 Summary of the Scientific Results of the Projects Supported within the Sonderforschungsbereich 'Molekularbiologie der Zelle'
2.5.3 'What was Discovered in the Fields Supported by the Sonderforschungsbereich?'
2.5.3.1 Metabolism of Membrane Lipids
2.5.3.2 Biophysical Studies on Membrane Lipids and Synthetic (Artificial) Bilayers4.13-22.31
2.5.3.3 High-Density Lipoproteins3.6.23-25.28.32
2.5.3.4 Molecular Neurobiology1.2.7.26.27.30
2.5.4 References
2.6 Resolution of Recombination Intermediates by Endonuclease VII in vitro
Borries Kemper
2.6.1 Summary
2.6.2 Preface
2.6.3 Introduction
2.6.4 Isolation and Characterization of VFS-DNA
2.6.5 Isolation and Characterization of Endonuclease VII
2.6.6 Endonuclease VII Resolves Holliday-Structures
2.6.7 Endonuclease VII Cleaves Secondary Structures in DNA
2.6.8 Endo VII Cleaves at Mismatches
2.6.9 Purification of Endonuclease X3 from Yeast Saccharomyces cerevisiae
2.6.10 Collaborations
2.6.11 The Model
2.6.12 References
3 Plant Molecular Biology
3.1 Transposable Elements in Bacteria and in Plants
Heinz Saedler and Peter Starlinger
3.1.1 Summary
3.1.2 The Discovery of IS-Elements
3.1.3 Studies on IS-Elements in Koln (H. Saedler)
3.1.4 Studies on IS-Elements in Koln (P. Starlinger)
3.1.5 Work on Transposable Element Ac (P. Starlinger)
3.1.6 Studies on Plant Molecular Biology in Koln (H. Saedler)
3.1.7 Open Questions
3.1.8 Publications
3.1.8.1 On the IS-Work
3.1.8.2 On the Work with Plants
3.2 Experiments to Analyze the Maize Transposable Elements Ac and Ds at the DNA Level
Hans-Peter Doring
3.2.1 Introduction
3.2.2 Isolation of the Chromosome-Breaking Ds-Element
3.2.3 How to Explain Chromosome Breakage at the Double Ds Structure
3.2.4 Chromosome-Breaking Ds-Elements can Generate Complicated Chromosomal Rearrangements
3.2.5 How was the Symmetrical Insert in the sh-m5933 Allele Created?
3.2.6 References
3.2.7 Publications and Review Articles
3.3 Agrobacterium - Mediated Gene Transfer to Plants: Its Impact on Plant Breeding, Studies of Gene Expression and Plant Developmental Biology
Jeff Schell
3.3.1 Introduction
3.3.2 Plant Developmental Biology
3.3.2.1 Identification, Cloning, and Characterization of Plant Genes Involved in Signal Transduction
3.3.2.2 Elucidation of the Mechanism of Action of Oncogenic T-DNA Genes from A. rhizogenes
3.3.2.3 Identification of Genes Involved in Control of Plant Development by Gene-Tagging Mutagenesis
3.3.3 Regulation of Gene Expression in Plants
3.3.3.1 Light Inducible Genes
3.3.3.2 Organ-Specific Genes
3.3.3.3 Stress Induced Genes
3.3.4 Transformation of Cereal Plants
3.3.5 The Rhizobium-Legume Symbiosis as a Model System for Plant-Bacterial Interactions
3.3.6 Molecular Biology and Potential Application of DNA Plant Viruses
3.3.7 Projects with Long Term Applied Goal
3.3.8 Publications
3.4 Resistance Mechanisms in Plants
Klaus Hahlbrock
3.4.1 Summary
3.4.2 Nonhost Disease Resistance in Parsley
3.4.3 UV Resistance in Parsley
3.4.4 Host Disease Resistance in Potato
3.4.5 Interaction of Barley with Rhynchosporium
3.4.6 Arabidopsis
4 Development and Differentiation
4.1 Neurogenesis in Drosophila
Jose Antonio Campos- Ortega
4.1.1 Introduction
4.1.2 Cellular Aspects of Neurogenesis
4.1.2.1 The Proliferation of the Neuroblasts
4.1.2.2 Cell Commitment in the Neurogenic Ectoderm
4.1.3 Genetics of Neurogenesis
4.1.3.1 The Neurogenic Genes
4.1.3.2 Functional Interactions Between the Neurogenic Loci
4.1.3.3 Genetic Organization of the Subdivision 1B
4.1.3.4 The Genetics and Molecular Genetics of the Daughterless Locus
4.1.3.5 Interactions Between Neurogenic and Genes of the AS-C and da
4.1.4 References
4.1.5 Publications from 1985 to 1988
4.2 The Function of Specific Nuclear Proteins in the Regulation of Gene Expression in Drosophila
Harald Saumweber
4.2.1 Introduction
4.2.2 Puff Specific Proteins
4.2.2.1 Mapping the Binding Sites Bj6 and Bx42 Proteins in situ
4.2.2.2 Structure and Expression of the Bj6 Gene
4.2.2.3 Isolation of Bj6 Mutations
4.2.3 Puff Specific Proteins: Structure and Expression of the Bx42 Gene
4.2.4 The Telomere Associated Protein At5
4.2.5 References
4.2.6 Publications from 1986 to 1988
4.3 Differentiation, Pattern Formation, and Sexual Induction in Volvox carteri
Lothar Jaenicke and Rainer Gilles
4.3.1 Introduction
4.3.2 Morphological Basis
4.3.3 Sexc-Switch
4.3.4 Sex Inducer
4.3.4.1 Chemical Structure
4.3.4.2 Membrane Receptor
4.3.5 Inducer Action
4.3.6 cAMP Signal Chain
4.3.6.1 Adenylate Cyclase and Cyclic Phosphodiesterase
4.3.6.2 Protein Kinases and cAMP Binding Protein
4.3.7 Calcium and Calmodulin
4.3.8 Conclusion
4.3.9 References
5 Immunology
5.1 Selection of the Receptor Repertoire in the Immune System
Klaus Rajewsky
5.1.1 The Starting Point: T-B Cell Collaboration
5.1.2 Interaction of (Helper) T and (Antibody Producing) B Cells
5.1.2.1 Helper Cell Specificity
5.1.2.2 Memory and Immunological Tolerance in T and B Cells
5.1.2.3 Mechanism of T-B Cell Interaction
5.1.3 Network Regulation
5.1.3.1 Idiotypic Research Gets Underway
5.1.3.2 Idiotype Suppression and the Problem of Network-Controlled Tolerance
5.1.3.3 Idiotype Stimulation and Anti-Idiotypic Vaccination
5.1.3.4 Unsuccessful Attempts to Solve the T Cell Receptor Problem
5.1.3.5 'Direct' Idiotypic T-B Cell Interaction
5.1.3.6 Structural Analysis of the Idiotypic Network
5.1.4 B Cell Differentiation and the Selection of the Antibody Repertoire
5.1.4.1 Early Differentiation Processes
5.1.4.2 Antibody Class Switching
5.1.4.3 Antibody V Regions: Selection and Somatic Diversification
5.1.5 Side Projects
5.1.6 References
5.2 Immunochemical Analysis of Antibody Binding Sites, Surface Immunoglobulins and Anti-Peptide Antibodies
Matthias Cramer
5.2.1 Summary
5.2.2 Structural Analysis of the Antibody Binding Site by Photoaffinity Labelling
5.2.3 The Use of Anti-Peptide Antibodies in the Study of Alzheimer's Disease
5.2.4 The Association of Beta-2-Microglobulin to MHC Antigens
5.2.5 Anchoring Surface Immunoglobulin in the Membrane
5.2.6 Cooperations
5.2.7 References
5.3 Germinal Centres and the Maturation of the Immune Response
Claudia Berek
5.3.1 The Immune Response to 2-Phenyl-oxazolone
5.3.2 Does the Micro-Environment of the Germinal Centre Play a Role in the Maturation of the Immune Response?
5.3.3 Analysis of Germinal Centre B Cells
5.3.3.1 Isolation of PNAhi and PNAlo B Cells
5.3.3.2 Fusions of PNAhi and PNAlo Cells
5.3.3.3 Antibody Diversity in Germinal Centre B Cells
5.3.4 Conclusions
5.3.5 Future
5.3.6 References
5.3.7 Publications
5.4 Genetic Basis of Germ-Line Encoded Antibody Diversity
Ulrich Krawinkel
5.4.1 Generation of V-Region Gene Diversity in the Germ-Line
5.4.1.1 Multiplicity and Diversity of VH-Genes in the Mouse
5.4.1.2 Organization of the Igh V Locus in Mice
5.4.2 Class Switch Recombination
5.4.3 References
6 Virology
6.1 Malignant Transformation by Adenovirus: Integration of the Viral DNA and Control of Viral Gene Expression by Specific Patterns of DNA Methylation
Walter Doerfler
6.1.1 Setting the Goal
6.1.2 Continuing Research in Koln
6.1.3 Insertional Mutagenesis as a Model for Transformation by Oncodna Viruses
6.1.4 On the Mechanism of Integrative Recombination in Mammalian Cells
6.1.5 The Species-Specificity of the Major Late Promoter of Ad12 DNA
6.1.5.1 Ad12-Specific Functions in Hamster Cells
6.1.5.2 A Mitigator Element in the Ad12 MLP
6.1.5.3 Partial Complementation of Ad12 Defects in Hamster Cells by Ad2 and Ad5 Functions
6.1.6 The Long-Term Inhibition of Promoter Activity by Sequence-Specific Methylation
6.1.6.1 Inverse Correlations
6.1.6.2 In vitro Methylation of Promoter Sequences and Effect on Activity
6.1.6.3 Reversal of Promoter Inactivation
6.1.6.4 The Spreading of DNA Methylation
6.1.6.5 Sequence-Specific Methylations and DNA-Protein Interactions in Eukaryotic Promoters
6.1.7 Transcription of the Genome of Autographa californica Nuclear Polyhedrosis Virus (Baculovirus)
6.1.8 References
6.2 Adenovirus DNA: Structure and Function of a Novel Replicon
Ernst-L. Winnacker
6.2.1 Introduction
6.2.2 Viral and Host Functions in the Lytic Infection Cycle
6.2.3 Initiation of Adenovirus DNA Replication
6.2.4 Summary
6.2.5 References
6.3 Malignant Transformation by Adenoviruses: Early Viral Gene Expression and Host Cell Response
Helmut Esche
6.3.1 Introduction
6.3.2 Summaries
6.3.2.1 Structure and Expression of Early Viral Gene Products in Adenovirus Type 12-Infected Human (KB, HELA) and -Transformed Rodent (Hamster, Mouse, Rat) Cells
6.3.2.2 Cell Factor(s) Mediated DNA-Binding of an Adenovirus Type 12 Coded Tumor-Antigen
6.3.2.3 Interaction of Adenovirus Gene Functions with the Regulation of the Host Cell Cycle
6.3.3 References
6.3.4 Publications
6.4 Replication of an Alphavirus in Cultured Vertebrate and Insect Cells
Dennis T. Brown
6.4.1 Introduction
6.4.2 The Development of Sindbis Virus in Tissue-Cultured Vertebrate Cells
6.4.3 Replication of Sindbis Virus in Cultured Insect Cells
6.4.4 References
6.4.5 Publications
7 Gene Transfer
Chromosome Mediated Transfer of Human Genes
Klaus Willecke
Biographical Notes