Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling:
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| Beschreibung: | Heidelberg, Univ., Diss., 2007 |
| Beschreibung: | XI, 163 S. graph. Darst. |
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| 245 | 1 | 0 | |a Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling |c [Celine Maeder] |
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| adam_text | Table of content
ACKNOWLEDGEMENTS V
CONTRIBUTIONS THROUGH COLLABORATIONS VII
1 SUMMARY 1
1 ZUSAMMENFASSUNG 5
2 INTRODUCTION 9
2.1 Mitogen activated protein kinase (MAPK) signalling 11
2.1.1 A general overview 11
2.1.2 MAPK signalling in S.cerevisiae 12
2.1.3 Pheromone signalling as a paradigmatic MAPK pathway in yeast 13
2.1.3.1 Pheromone induced mating 13
2.1.3.2 Overview of the pheromone response pathway 15
2.1.3.3 Regulation of the MAPK scaffold module 18
2.1.3.3.1 Regulation of Ste 11 18
2.1.3.3.2 Regulation of Ste7 20
2.1.3.3.3 The various roles of Ste5 in pheromone signalling 21
2.1.3.3.3.1 Ste5 as an assembly platform for signalling molecules 21
2.1.3.3.3.2 Ste5 and its functions as a scaffold 21
2.1.3.3.3.3 Ste5 oligomerization is essential for signalling 22
2.1.3.3.3.4 Dynamic Ste5 localization in pheromone signalling 23
2.1.3.3.3.5 Cell cycle regulation of Ste5 activity 24
2.1.3.3.3.6 Ste5 allosterically modulates signalling output 25
2.1.3.3.4 The diverse effector roles of Fus3 in pheromone signalling 25
2.1.3.3.4.1 Fus3 and its role in signal attenuation 26
2.1.3.3.4.2 Fus3 and its role in transcriptional regulation of the
pheromone response 26
2.1.3.3.4.3 Fus3 and its role in cell cycle control 27
2.1.3.3.4.4 Fus3 and its role in cell polarization 28
2.1.3.3.4.5 Fus3 and its role in pathway specificity 29
2.1.3.3.4.6 Fus3 activity regulation by phosphatases 30
2.2 Fluorescence correlation and cross correlation spectroscopy 31
2.2.1 Fluorescence correlation spectroscopy 31
2.2.1.1 Qualitative description of FCS and the autocorrelation function 33
2.2.1.2 Mathematical description of the autocorrelation curve 34
2.2.2 Fluorescence cross correlation spectroscopy 37
2.2.2.1 A qualitative description of FCCS and the cross correlation curve 37
2.2.2.2 Mathematical description of the cross correlation curve 39
2.2.3 Photon counting histogram 40
I
Table of content
2.2.4 Considerations and limitations of FCS/FCCS 41
Dynamic range and photobleaching 41
Signal to noise ratio, background fluorescence and
molecular brightness 42
Crosstalk 42
Incomplete observation volume overlap 43
2.3 Aim of the project 43
3 RESULTS 45
3.1 FCS/FCCS establishment in yeast 47
3.1.1 Triple tags of fluorescent proteins yield a high molecular brightness
suitable for FCS/FCCS experiments 47
3.1.2 Choosing the optimal partner for GFP in FCCS experiments: a walk
through the red fluorescent proteins 49
3.1.3 Red fluorescent proteins show a maturation delay in yeast cells 50
3.1.4 How reliable are quantitative FCS measurements: a comparison to
quantitative Western blotting 53
3.1.5 FCCS as a valid method to detect complexes in living yeast cells 55
3.2 Quantitative analysis of MAPK scaffold complexes in the yeast
pheromone signalling cascade 56
3.2.1 Components of the MAPK module fused to fluorescent tags are
functional 56
3.2.2 Localization of the MAPK module under endogenous expression
conditions 58
3.2.3 Concentration determination of the MAPK module in vegetative and
stimulated cells by FCS 59
3.2.4 Pheromone stimulation does not change cytoplasmic MAPK complexes...61
3.2.4.1 Concentration analysis of the complexes between components the
MAPK scaffold module by FCCS 61
3.2.4.2 KJf analysis of the complexes between components the MAPK
scaffold module by FCCS 63
3.2.5 Ste5 does not dimerize in the cytoplasm of vegetative and pheromone
stimulated cells 66
3.2.6 Binary and higher order interactions between cytoplasmic MAP kinase
scaffold components 67
3.2.6.1 Binary and higher order interactions between the components of the
MAPK scaffold module 68
3.2.6.2 Deletion of Ste5 does not affect complex formation between Fus3 with
Ste7 or Stel 1 71
3.2.6.3 Ste5 and Ste7 do not bind competitively to Fus3 under endogenous
conditions in living cells 73
3.2.6.4 Kssl binds strongly and not competitively to Ste7, but does not bind to
Ste5 75
Table of content
3.3 Recruitment of MAP kinases to the shmoo tip 77
3.3.1 Establishment of quantitative photon counting imaging 77
3.3.2 High enrichment of Fus3 and Ste7, relative to Ste5 at the shmoo tip 78
3.3.3 Does Spa2 serve as scaffold for Ste7 recruitment to the shmoo? 80
3.3.4 Fus3 enrichment does not solely depend on Ste5 and pathway activity 82
3.3.5 Fus3 enrichment in the shmoo via enzyme substrate interactions 83
3.3.6 Cytoplasmic phosphatase levels influence Fus3 localization to the shmoo.85
3.4 A cytoplasmic gradient of active Fus3lp emanating from the shmoo tip..88
3.4.1 Mathematical modelling of an active Fus3pp gradient 88
3.4.2 Visualization of the active Fus3pp gradient by FL1M (fluorescence life
time imaging) 89
i 4 DISCUSSION 95
4.1 Advantages and limitations of FCCS in yeast 98
4.2 Pheromone induced MAPK signalling is not regulated by changes in
cytoplasmic complexes 100
4.3 Pheromone signalling is not mediated via Ste5 dimerization in the
cytoplasm 101
4.4 Components of the MAPK scaffold module interact through
equilibrium binding 102
4.4.1 Fus3 Ste7 complexes are very stable, mediated via direct binding and do not
show any competitive behaviour with Kssl and Ste5 103
4.4.2 Components of the MAPK scaffold module bind cooperatively to form
macromolecular complexes 104
4.5 Ste5 dependent and independent mechanisms account for the shmoo tip
localization of Ste7 106
4.6 Fus3 accumulation at the shmoo tip due to enzyme substrate
interactions 108
4.7 Cytoplasmic phosphatases and shmoo tip localized activating kinases
generate a gradient of active Fus3pr throughout the cell 110
4.8 What might be the role of a gradient of active Fus3pp in pheromone
signalling? 113
i
4.8.1 A role in modulation of transcriptional regulation? 113
: 4.8.2 A role in nuclear positioning within the cell? 114
i 4.8.3 A role in endocytosis to establish and maintain accurate polarization?.... 115
i III
Table of content
5 MATERIALS METHODS 117
5.1 Materials 119
5.2 Methods 122
5.2.1 Yeast methods 122
5.2.1.1 Growth media and growth conditions 122
5.2.1.2 PCR targeting of yeast genes 122
5.2.1.3 Replacement of wild type gene with mutant alleles 123
5.2.1.4 Fluorescent protein constructs 123
5.2.1.5 Whole gene synthesis of dTomato 124
5.2.1.6 Antibody production and purification 124
5.2.1.7 Trichloroacetic acid cell lysis of whole yeast cells 125
5.2.1.8 SDS PAGE 125
5.2.1.9 Phos tag SDS PAGE 125
5.2.1.10 Western blotting 126
5.2.1.11 Functionality assays 127
5.2.1.11.1 Halo assay 127
5.2.1.11.2 Transcriptional response assay 127
5.2.2 Microscopy 128
5.2.2.1 Sample perparation 128
5.2.2.2 Confocal photon counting (APD) imaging 128
5.2.2.3 Quantitative image analysis 129
5.2.2.4 FCS/FCCS 130
5.2.2.4.1 Data acquisition 130
5.2.2.4.2 Data analysis 131
5.2.2.5 Fluorescence lifetime imaging microscopy (FLIM) 133
5.2.2.5.1 Sample preparation 133
5.2.2.5.2 Image acquisition and analysis 134
5.2.3 Mathematical analysis and modelling 135
5.2.3.1 Log normal distributions 135
5.2.3.2 Protein interaction analysis 136
5.2.3.2.1 The binary dissociation constant KD (KD) 136
5.2.3.2.2 The effective KD(Kjf) 136
5.2.3.3 Reaction diffusion model of active Fus3 gradient 138
6 APPENDIX 143
6.1 Abbreviations 145
6.2 List of tables 146
6.3 List of figures 146
7 REFERENCES 149
T1 7
6.2 List of tables
Table 1: Statistical analysis of FCS and FCCS data 65
Table 2: Genotypes of yeast strains 119
Table 3: Genotypes of plasmids 121
6.3 List of figures
Figure 1: MAPK signalling in S.cerevisiae 14
Figure 2: Pheromone induced mating in S.cerevisiae. 16
Figure 3: MAPK signalling upon pheromone stimulation 18
Figure 4: Schematic representation of Ste5 domains 21
Figure 5: Ste5 localization to the plasma membrane is only facilitated when it
interacts with lipids and Gbg 24
Figure 6: Confocal microscopy setup for FCS and FCCS measurements 31
Figure 7: Schematic representation of FCS 32
Figure 8: Qualitative representation of the autocorrelation curve 34
Figure 9: Principal of FCCS 38
Figure 10: Qualitative representation of the cross correlation curve 38
Figure 11: Molecular brightness analysis of multiple meGFP tags 49
Figure 12: Characteristics of red fluorescent proteins 50
Figure 13: Characterization of red fluorescent proteins 52
Figure 14: Quantitative Western blotting 54
Figure 15: Validation of FCCS measurements in yeast using control
constructs 56
Figure 16: Expression of fluorescent protein tagged proteins does not interfere with
pheromone signalling 57
Figure 17: Localization of endogenously expressed Ste5, Ste7, Stel 1 and Fus3 in
vegetative and pheromone stimulated cells 59
Figure 18: Concentration determination of the MAPK module in the cytoplasm of
vegetative and stimulated cells 60
Figure 19: Quantification of complexes between Ste5, Ste7, Stel 1 and Fus3 in the
cytoplasm 62
Figure 20: FCS/FCCS measurements are independent on the specific fluorescent
protein used for gene fusion 63
Figure 21: Ste5 does not dimerize in the cytoplasm of vegetative and pheromone
stimulated cells 67
Figure 22: Mathematical simulation of the effect of neutral, cooperative and
competitive binding on the interaction of two components 68
Figure 23: Statistical analysis of the binding isotherms of complexes formed
between pairs of MAPK fusion proteins 69
Figure 24: Deletion of Ste5 does not affect complex formation between Fus3 and
Ste7 / Stel 1 72
Figure 25 Ste5 and Ste7 do not competitively bind to Fus3 under endogenous
conditions 74
Figure 26: Interaction of Kssl with Ste5 and Ste7 and its localization 76
6. Appendix
Figure 27: Proof of principle for the quantitative measurement of protein
abundance in the shmoo using FCS data for the calibration of
quantitative APD imaging 78
Figure 28: Quantification of relative MAP kinase abundances at different sites in
yeast cells 79
Figure 29: Spa2 interacts with Ste7 and does not affect Ste7 Stel 1 complex 81
Figure 30: Fus3 localization to the shmoo does not largely depend on Ste5 83
Figure 31: Fus3 enrichment in the shmoo via enzyme substrate interactions 85
Figure 32: Cytoplasmic phosphatase levels influence the localization of Fus3 to
the shmoo 87
Figure 33: Computational analysis of an active Fus3pp gradient 89
Figure 34: Visualization of the active Fus3pp gradient in stimulated cells 91
Figure 35: Overview about all possible complexes of the MAPK scaffold module.
101
Figure 36: Current model of the pheromone induced MAPK signalling cascade
for vegetative 112
Figure 37: Model for cell polarization and nuclear positioning in fully
differentiated cells 116
Figure 38: Comparison of normal and lognormal distributions 135
|
| adam_txt |
Table of content
ACKNOWLEDGEMENTS V
CONTRIBUTIONS THROUGH COLLABORATIONS VII
1 SUMMARY 1
1 ZUSAMMENFASSUNG 5
2 INTRODUCTION 9
2.1 Mitogen activated protein kinase (MAPK) signalling 11
2.1.1 A general overview 11
2.1.2 MAPK signalling in S.cerevisiae 12
2.1.3 Pheromone signalling as a paradigmatic MAPK pathway in yeast 13
2.1.3.1 Pheromone induced mating 13
2.1.3.2 Overview of the pheromone response pathway 15
2.1.3.3 Regulation of the MAPK scaffold module 18
2.1.3.3.1 Regulation of Ste 11 18
2.1.3.3.2 Regulation of Ste7 20
2.1.3.3.3 The various roles of Ste5 in pheromone signalling 21
2.1.3.3.3.1 Ste5 as an assembly platform for signalling molecules 21
2.1.3.3.3.2 Ste5 and its functions as a scaffold 21
2.1.3.3.3.3 Ste5 oligomerization is essential for signalling 22
2.1.3.3.3.4 Dynamic Ste5 localization in pheromone signalling 23
2.1.3.3.3.5 Cell cycle regulation of Ste5 activity 24
2.1.3.3.3.6 Ste5 allosterically modulates signalling output 25
2.1.3.3.4 The diverse effector roles of Fus3 in pheromone signalling 25
2.1.3.3.4.1 Fus3 and its role in signal attenuation 26
2.1.3.3.4.2 Fus3 and its role in transcriptional regulation of the
pheromone response 26
2.1.3.3.4.3 Fus3 and its role in cell cycle control 27
2.1.3.3.4.4 Fus3 and its role in cell polarization 28
2.1.3.3.4.5 Fus3 and its role in pathway specificity 29
2.1.3.3.4.6 Fus3 activity regulation by phosphatases 30
2.2 Fluorescence correlation and cross correlation spectroscopy 31
2.2.1 Fluorescence correlation spectroscopy 31
2.2.1.1 Qualitative description of FCS and the autocorrelation function 33
2.2.1.2 Mathematical description of the autocorrelation curve 34
2.2.2 Fluorescence cross correlation spectroscopy 37
2.2.2.1 A qualitative description of FCCS and the cross correlation curve 37
2.2.2.2 Mathematical description of the cross correlation curve 39
2.2.3 Photon counting histogram 40
I
Table of content
2.2.4 Considerations and limitations of FCS/FCCS 41
Dynamic range and photobleaching 41
Signal to noise ratio, background fluorescence and
molecular brightness 42
Crosstalk 42
Incomplete observation volume overlap 43
2.3 Aim of the project 43
3 RESULTS 45
3.1 FCS/FCCS establishment in yeast 47
3.1.1 Triple tags of fluorescent proteins yield a high molecular brightness
suitable for FCS/FCCS experiments 47
3.1.2 Choosing the optimal partner for GFP in FCCS experiments: a walk
through the red fluorescent proteins 49
3.1.3 Red fluorescent proteins show a maturation delay in yeast cells 50
3.1.4 How reliable are quantitative FCS measurements: a comparison to
quantitative Western blotting 53
3.1.5 FCCS as a valid method to detect complexes in living yeast cells 55
3.2 Quantitative analysis of MAPK scaffold complexes in the yeast
pheromone signalling cascade 56
3.2.1 Components of the MAPK module fused to fluorescent tags are
functional 56
3.2.2 Localization of the MAPK module under endogenous expression
conditions 58
3.2.3 Concentration determination of the MAPK module in vegetative and
stimulated cells by FCS 59
3.2.4 Pheromone stimulation does not change cytoplasmic MAPK complexes.61
3.2.4.1 Concentration analysis of the complexes between components the
MAPK scaffold module by FCCS 61
3.2.4.2 KJf analysis of the complexes between components the MAPK
scaffold module by FCCS 63
3.2.5 Ste5 does not dimerize in the cytoplasm of vegetative and pheromone
stimulated cells 66
3.2.6 Binary and higher order interactions between cytoplasmic MAP kinase
scaffold components 67
3.2.6.1 Binary and higher order interactions between the components of the
MAPK scaffold module 68
3.2.6.2 Deletion of Ste5 does not affect complex formation between Fus3 with
Ste7 or Stel 1 71
3.2.6.3 Ste5 and Ste7 do not bind competitively to Fus3 under endogenous
conditions in living cells 73
3.2.6.4 Kssl binds strongly and not competitively to Ste7, but does not bind to
Ste5 75
Table of content
3.3 Recruitment of MAP kinases to the shmoo tip 77
3.3.1 Establishment of quantitative photon counting imaging 77
3.3.2 High enrichment of Fus3 and Ste7, relative to Ste5 at the shmoo tip 78
3.3.3 Does Spa2 serve as scaffold for Ste7 recruitment to the shmoo? 80
3.3.4 Fus3 enrichment does not solely depend on Ste5 and pathway activity 82
3.3.5 Fus3 enrichment in the shmoo via enzyme substrate interactions 83
3.3.6 Cytoplasmic phosphatase levels influence Fus3 localization to the shmoo.85
3.4 A cytoplasmic gradient of active Fus3lp emanating from the shmoo tip.88
3.4.1 Mathematical modelling of an active Fus3pp gradient 88
3.4.2 Visualization of the active Fus3pp gradient by FL1M (fluorescence life
time imaging) 89
i 4 DISCUSSION 95
4.1 Advantages and limitations of FCCS in yeast 98
4.2 Pheromone induced MAPK signalling is not regulated by changes in
cytoplasmic complexes 100
4.3 Pheromone signalling is not mediated via Ste5 dimerization in the
cytoplasm 101
4.4 Components of the MAPK scaffold module interact through
equilibrium binding 102
4.4.1 Fus3 Ste7 complexes are very stable, mediated via direct binding and do not
show any competitive behaviour with Kssl and Ste5 103
4.4.2 Components of the MAPK scaffold module bind cooperatively to form
macromolecular complexes 104
4.5 Ste5 dependent and independent mechanisms account for the shmoo tip
localization of Ste7 106
4.6 Fus3 accumulation at the shmoo tip due to enzyme substrate
interactions 108
4.7 Cytoplasmic phosphatases and shmoo tip localized activating kinases
generate a gradient of active Fus3pr throughout the cell 110
4.8 What might be the role of a gradient of active Fus3pp in pheromone
signalling? 113
i
4.8.1 A role in modulation of transcriptional regulation? 113
: 4.8.2 A role in nuclear positioning within the cell? 114
i 4.8.3 A role in endocytosis to establish and maintain accurate polarization?. 115
i III
Table of content
5 MATERIALS METHODS 117
5.1 Materials 119
5.2 Methods 122
5.2.1 Yeast methods 122
5.2.1.1 Growth media and growth conditions 122
5.2.1.2 PCR targeting of yeast genes 122
5.2.1.3 Replacement of wild type gene with mutant alleles 123
5.2.1.4 Fluorescent protein constructs 123
5.2.1.5 Whole gene synthesis of dTomato 124
5.2.1.6 Antibody production and purification 124
5.2.1.7 Trichloroacetic acid cell lysis of whole yeast cells 125
5.2.1.8 SDS PAGE 125
5.2.1.9 Phos tag SDS PAGE 125
5.2.1.10 Western blotting 126
5.2.1.11 Functionality assays 127
5.2.1.11.1 Halo assay 127
5.2.1.11.2 Transcriptional response assay 127
5.2.2 Microscopy 128
5.2.2.1 Sample perparation 128
5.2.2.2 Confocal photon counting (APD) imaging 128
5.2.2.3 Quantitative image analysis 129
5.2.2.4 FCS/FCCS 130
5.2.2.4.1 Data acquisition 130
5.2.2.4.2 Data analysis 131
5.2.2.5 Fluorescence lifetime imaging microscopy (FLIM) 133
5.2.2.5.1 Sample preparation 133
5.2.2.5.2 Image acquisition and analysis 134
5.2.3 Mathematical analysis and modelling 135
5.2.3.1 Log normal distributions 135
5.2.3.2 Protein interaction analysis 136
5.2.3.2.1 The binary dissociation constant KD (KD) 136
5.2.3.2.2 The effective KD(Kjf) 136
5.2.3.3 Reaction diffusion model of active Fus3 gradient 138
6 APPENDIX 143
6.1 Abbreviations 145
6.2 List of tables 146
6.3 List of figures 146
7 REFERENCES 149
T1 7
6.2 List of tables
Table 1: Statistical analysis of FCS and FCCS data 65
Table 2: Genotypes of yeast strains 119
Table 3: Genotypes of plasmids 121
6.3 List of figures
Figure 1: MAPK signalling in S.cerevisiae 14
Figure 2: Pheromone induced mating in S.cerevisiae. 16
Figure 3: MAPK signalling upon pheromone stimulation 18
Figure 4: Schematic representation of Ste5 domains 21
Figure 5: Ste5 localization to the plasma membrane is only facilitated when it
interacts with lipids and Gbg 24
Figure 6: Confocal microscopy setup for FCS and FCCS measurements 31
Figure 7: Schematic representation of FCS 32
Figure 8: Qualitative representation of the autocorrelation curve 34
Figure 9: Principal of FCCS 38
Figure 10: Qualitative representation of the cross correlation curve 38
Figure 11: Molecular brightness analysis of multiple meGFP tags 49
Figure 12: Characteristics of red fluorescent proteins 50
Figure 13: Characterization of red fluorescent proteins 52
Figure 14: Quantitative Western blotting 54
Figure 15: Validation of FCCS measurements in yeast using control
constructs 56
Figure 16: Expression of fluorescent protein tagged proteins does not interfere with
pheromone signalling 57
Figure 17: Localization of endogenously expressed Ste5, Ste7, Stel 1 and Fus3 in
vegetative and pheromone stimulated cells 59
Figure 18: Concentration determination of the MAPK module in the cytoplasm of
vegetative and stimulated cells 60
Figure 19: Quantification of complexes between Ste5, Ste7, Stel 1 and Fus3 in the
cytoplasm 62
Figure 20: FCS/FCCS measurements are independent on the specific fluorescent
protein used for gene fusion 63
Figure 21: Ste5 does not dimerize in the cytoplasm of vegetative and pheromone
stimulated cells 67
Figure 22: Mathematical simulation of the effect of neutral, cooperative and
competitive binding on the interaction of two components 68
Figure 23: Statistical analysis of the binding isotherms of complexes formed
between pairs of MAPK fusion proteins 69
Figure 24: Deletion of Ste5 does not affect complex formation between Fus3 and
Ste7 / Stel 1 72
Figure 25 Ste5 and Ste7 do not competitively bind to Fus3 under endogenous
conditions 74
Figure 26: Interaction of Kssl with Ste5 and Ste7 and its localization 76
6. Appendix
Figure 27: Proof of principle for the quantitative measurement of protein
abundance in the shmoo using FCS data for the calibration of
quantitative APD imaging 78
Figure 28: Quantification of relative MAP kinase abundances at different sites in
yeast cells 79
Figure 29: Spa2 interacts with Ste7 and does not affect Ste7 Stel 1 complex 81
Figure 30: Fus3 localization to the shmoo does not largely depend on Ste5 83
Figure 31: Fus3 enrichment in the shmoo via enzyme substrate interactions 85
Figure 32: Cytoplasmic phosphatase levels influence the localization of Fus3 to
the shmoo 87
Figure 33: Computational analysis of an active Fus3pp gradient 89
Figure 34: Visualization of the active Fus3pp gradient in stimulated cells 91
Figure 35: Overview about all possible complexes of the MAPK scaffold module.
101
Figure 36: Current model of the pheromone induced MAPK signalling cascade
for vegetative 112
Figure 37: Model for cell polarization and nuclear positioning in fully
differentiated cells 116
Figure 38: Comparison of normal and lognormal distributions 135 |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Maeder, Céline |
| author_GND | (DE-588)133345386 |
| author_facet | Maeder, Céline |
| author_role | aut |
| author_sort | Maeder, Céline |
| author_variant | c m cm |
| building | Verbundindex |
| bvnumber | BV022883241 |
| ctrlnum | (OCoLC)633076484 (DE-599)GBV546247903 |
| format | Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV022883241 |
| illustrated | Illustrated |
| index_date | 2024-07-02T18:51:12Z |
| indexdate | 2024-07-09T21:07:41Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-016088193 |
| oclc_num | 633076484 |
| open_access_boolean | |
| owner | DE-29T |
| owner_facet | DE-29T |
| physical | XI, 163 S. graph. Darst. |
| publishDate | 2006 |
| publishDateSearch | 2006 |
| publishDateSort | 2006 |
| record_format | marc |
| spelling | Maeder, Céline Verfasser (DE-588)133345386 aut Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling [Celine Maeder] 2006 XI, 163 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier Heidelberg, Univ., Diss., 2007 MAP-Kinase (DE-588)4576144-9 gnd rswk-swf Fluoreszenzkorrelationsspektroskopie (DE-588)4636290-3 gnd rswk-swf Saccharomyces cerevisiae (DE-588)4178812-6 gnd rswk-swf Pheromon (DE-588)4174258-8 gnd rswk-swf Multiproteinkomplex (DE-588)4798068-0 gnd rswk-swf Signalkette (DE-588)4409935-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content MAP-Kinase (DE-588)4576144-9 s Signalkette (DE-588)4409935-6 s Saccharomyces cerevisiae (DE-588)4178812-6 s Pheromon (DE-588)4174258-8 s Fluoreszenzkorrelationsspektroskopie (DE-588)4636290-3 s Multiproteinkomplex (DE-588)4798068-0 s DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016088193&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Maeder, Céline Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling MAP-Kinase (DE-588)4576144-9 gnd Fluoreszenzkorrelationsspektroskopie (DE-588)4636290-3 gnd Saccharomyces cerevisiae (DE-588)4178812-6 gnd Pheromon (DE-588)4174258-8 gnd Multiproteinkomplex (DE-588)4798068-0 gnd Signalkette (DE-588)4409935-6 gnd |
| subject_GND | (DE-588)4576144-9 (DE-588)4636290-3 (DE-588)4178812-6 (DE-588)4174258-8 (DE-588)4798068-0 (DE-588)4409935-6 (DE-588)4113937-9 |
| title | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling |
| title_auth | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling |
| title_exact_search | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling |
| title_exact_search_txtP | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling |
| title_full | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling [Celine Maeder] |
| title_fullStr | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling [Celine Maeder] |
| title_full_unstemmed | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling [Celine Maeder] |
| title_short | Quantitative imaging of MAPK-Scaffold complexes reveals spatial regulation of Fus3 activity in Yeast Pheromone Signalling |
| title_sort | quantitative imaging of mapk scaffold complexes reveals spatial regulation of fus3 activity in yeast pheromone signalling |
| topic | MAP-Kinase (DE-588)4576144-9 gnd Fluoreszenzkorrelationsspektroskopie (DE-588)4636290-3 gnd Saccharomyces cerevisiae (DE-588)4178812-6 gnd Pheromon (DE-588)4174258-8 gnd Multiproteinkomplex (DE-588)4798068-0 gnd Signalkette (DE-588)4409935-6 gnd |
| topic_facet | MAP-Kinase Fluoreszenzkorrelationsspektroskopie Saccharomyces cerevisiae Pheromon Multiproteinkomplex Signalkette Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016088193&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT maederceline quantitativeimagingofmapkscaffoldcomplexesrevealsspatialregulationoffus3activityinyeastpheromonesignalling |