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High-Resolution Profiling of Protein-RNA Interactions 2015 ed. [Hardback]

  • Formāts: Hardback, 121 pages, height x width: 235x155 mm, weight: 3495 g, 20 Illustrations, color; 5 Illustrations, black and white; XXIII, 121 p. 25 illus., 20 illus. in color., 1 Hardback
  • Sērija : Springer Theses
  • Izdošanas datums: 27-Mar-2015
  • Izdevniecība: Springer International Publishing AG
  • ISBN-10: 3319162527
  • ISBN-13: 9783319162522
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  • Formāts: Hardback, 121 pages, height x width: 235x155 mm, weight: 3495 g, 20 Illustrations, color; 5 Illustrations, black and white; XXIII, 121 p. 25 illus., 20 illus. in color., 1 Hardback
  • Sērija : Springer Theses
  • Izdošanas datums: 27-Mar-2015
  • Izdevniecība: Springer International Publishing AG
  • ISBN-10: 3319162527
  • ISBN-13: 9783319162522
Citas grāmatas par šo tēmu:
The work reported in this book represents an excellent example of how creative experimentation and technology development, complemented by computational data analysis, can yield important insights that further our understanding of biological entities from a systems perspective. The book describes how the study of a single RNA-binding protein and its interaction sites led to the development of the novel "protein occupancy profiling" technology that for the first time captured the mRNA sequence space contacted by the ensemble of expressed RNA binders. Application of protein occupancy profiling to eukaryotic cells revealed that extensive sequence stretches in 3" UTRs can be contacted by RBPs and that evolutionary conservation as well as negative selection act on protein-RNA contact sites, suggesting functional importance. Comparative analysis of the RBP-bound sequence space has the potential to unravel putative cis -acting RNA elements without a priori knowledge of the bound regulato

rs. Here, Dr. Munschauer provides a comprehensive introduction to the field of post-transcriptional gene regulation, examines state-of-the-art technologies, and combines the conclusions from several journal articles into a coherent and logical story from the frontiers of systems-biology inspired life science. This thesis, submitted to the Department of Biology, Chemistry and Pharmacy at Freie Universität Berlin, was selected as outstanding work by the Berlin Institute for Medical Systems Biology at the Max-Delbrueck Center for Molecular Medicine, Germany.

Introduction.- Mapping Regulatory Interactions of the Rna-Binding Protein Lin28b.- Exploring The Sequence Space Contacted by the Ensemble of Rna-Binding Proteins.- Revealing Cell-Type Specific Differences in Protein Occupancy on Rna.- Discussion.
1 Introduction
1(48)
1.1 The Life Cycle of an Eukaryotic mRNA Molecule
1(4)
1.2 RNA-Binding Proteins
5(2)
1.3 RNA-Binding Proteins and Disease
7(3)
1.4 The RNA-Binding Protein LIN28
10(5)
1.4.1 Lin28 Inhibits miRNA Let-7 Biogenesis
11(1)
1.4.2 Mechanism of Lin28-Let-7 Recognition
12(1)
1.4.3 The Functional Role of Lin28 in Stem Cell Biology, Cancer and Metabolism
12(2)
1.4.4 Lin28 Can Function Independent of Let-7
14(1)
1.4.5 Lin28 as a Direct Regulator of Mrna Translation
14(1)
1.5 MicroRNAs
15(3)
1.6 Cis-regulatory Sequence Elements in Eukaryotes
18(9)
1.6.1 Upstream Open Reading Frames (uORFs)
18(1)
1.6.2 Internal Ribosome Entry Sites (IRESs)
19(1)
1.6.3 Ribosome Frameshift Signals (RFSs)
20(1)
1.6.4 Splicing Regulatory Elements (SREs)
21(1)
1.6.5 Iron Response Elements (IREs)
22(1)
1.6.6 RNA Methylation Sites
22(2)
1.6.7 AU-Rich Elements (AREs)
24(1)
1.6.8 Zipcodes
25(1)
1.6.9 Polyadenylation Signals (PASs)
26(1)
1.7 Target Site Identification of Post-transcriptional Regulators
27(22)
1.7.1 From the Study of a Single RBP to the `Post-transcriptional Regulatome'
32(1)
References
33(16)
2 Mapping Regulatory Interactions of the RNA-Binding Protein LIN28B
49(12)
2.1 PAR-CLIP Reproducibly Identifies Thousands of Human RNAs Directly Bound by LIN28B
49(2)
2.2 LIN28B Binds to Let-7 Precursors and Protein Coding Transcripts
51(2)
2.3 Target Transcripts Are Enriched for a RGGSWG Consensus Motif
53(1)
2.4 Individual Domain PAR-CLIP Enables Characterization of Domain Specific Target Interactions
54(2)
2.5 LIN28B Enhances Protein Production of mRNA Target Transcripts
56(2)
2.6 LIN28B Controls Core Cell Cycle Regulators
58(3)
References
59(2)
3 Exploring the Sequence Space Contacted by the Ensemble of RNA-Binding Proteins
61(12)
3.1 Protein Occupancy Profiling Provides Catalog of Protein-mRNA Contact Sites
62(2)
3.2 Protein Occupancy Profiling Recapitulates AGO Binding Pattern at miRNA Target Sites
64(2)
3.3 Protein Occupancy Profiling Reveals Widespread and Conserved Protein-mRNA Contacts
66(2)
3.4 Putative RNA Cis-regulatory Elements Overlap with Trait/Disease-Associated Polymorphisms
68(1)
3.5 The Impact of Actively Translating Ribosomes on Protein Occupancy Profiles
68(5)
References
71(2)
4 Revealing Cell-Type Specific Differences in Protein Occupancy on RNA
73(16)
4.1 Protein Occupancy Profiling in MCF7 Cells
73(1)
4.2 Comparing Gene Expression and Protein Occupancy Profiles in MCF7 and HEK293 Cells
74(3)
4.3 Differential Protein Occupancy Profiling Based on T-C Transitions
77(1)
4.4 Identification of Differentially Occupied RNA Regions Between MCF7 and HEK293 Cells
78(3)
4.5 Differentially Occupied Positions Show Distinct Secondary-Structure Characteristics and Overlap with Binding Sites of Known RBPs
81(4)
4.6 Transcripts with Increased Protein Occupancy in MCF7 Cells Show Elevated mRNA Half-Lives
85(4)
References
86(3)
5 Discussion
89(32)
5.1 PAR-CLIP and iDo-PAR-CLIP: Challenges and Considerations
90(3)
5.1.1 PAR-CLIP Depends on Effective Metabolic Labeling of RNA
90(1)
5.1.2 Potential Biases in CLIP and PAR-CLIP Experiments
91(1)
5.1.3 Individual Domain PAR-CLIP: Asymmetry Is Key
92(1)
5.2 Controlling Background in CLIP and PAR-CLIP Experiments
93(2)
5.2.1 The Advantage of Combining Old and New Ways to Capturing RNA Targets
93(1)
5.2.2 The Challenge of the Next Generation: Controlling Sequencing Depth
94(1)
5.3 Transcriptome-Wide Identification of LLN28B-Bound RNA Targets
95(6)
5.3.1 Multiple Studies Identify Lin28A and Lin28B-Bound RNA Targets in Different Systems
95(2)
5.3.2 From Transcriptome-Wide Lin28 Binding Sites to a Model of mRNA Recognition
97(2)
5.3.3 A Direct Role for LIN28B in Regulating Protein Synthesis
99(2)
5.4 The Emerging Picture: Protein Production Is Regulated by Lin28 Through Let-7-Dependent and Let-7-Independent Mechanisms
101(2)
5.4.1 Let-7-Dependent Effects of Lin28
101(1)
5.4.2 Let-7-Independent Effects of Lin28
102(1)
5.4.3 Merging Two Worlds: MRNA Translation Is Directly Regulated by Lin28 and Let-7
102(1)
5.5 Transcriptome-Wide Protein Occupancy Profiling
103(18)
5.5.1 Protein Occupancy Profiling and the mRNA Bound Proteome
104(1)
5.5.2 Characteristics of Protein Occupancy Profiles
105(2)
5.5.3 Protein Occupancy and mRNA-Expression: A Distant Relationship?
107(1)
5.5.4 Protein Occupancy and Ribosomes: An Unexpected Crosslinking Bias
107(2)
5.5.5 Differential Protein Occupancy: From Crosslinks to Regulators
109(2)
5.5.6 Transcriptome-Wide and Unbiased Identification of Novel Cis-acting RNA Elements
111(1)
5.5.7 Lessons from the RBP-Bound mRNA Sequence Space
111(1)
5.5.8 Application of Protein Occupancy Profiling and Future Directions
112(2)
References
114(7)
Supplementary Information 121