Time-lapse Microscopy and Fluorescence Resonance Energy Transfer to Analyze the Dynamics and Interactions of Nucleolar Proteins
The dynamics of proteins play a key role in the organization and control of nuclear functions. Techniques were developed recently to observe the movement and interactions of proteins in living cells; time-lapse microscopy using fluorescent-tagged proteins gives access to observations of nuclear protein trafficking over time, and fluorescence resonance energy transfer (FRET) is used to investigate protein interactions in the time-lapse mode. In this chapter, we describe the application of these two approaches to follow the recruitment of nucleolar processing proteins at the time of nucleolar assembly. We question the role of prenucleolar bodies (PNB) during migration of the processing proteins from the chromosome periphery to sites of ribosomal genes (rDNA) transcription. The order of recruitment of different processing proteins into nucleoli is the consequence of differential sorting from the same PNBs. The dynamics of the interactions between processing proteins in PNBs suggest that PNBs are preassembly platforms for ribosomal RNA (rRNA) processing complexes.
- High Performance Liquid Chromatography/Electron Spin Resonance/Mass Spectrometry Analyses of Lipid-Derived Radicals
- Experimental Validation of MicroRNA Targets Using a Luciferase Reporter System
- Convergent Extension Analysis in Mouse Whole Embryo Culture
- RC4DRestriction Fragment Length Polymorphism-Coupled Domain-Directed Differential Display
- Animal Models of Multiple Sclerosis
- Ultrastructural and Immunofluorescent Methods for the Study of the XY Body as a Biomarker
- Screening Mutant Libraries in Saccharomyces cerevisiae
- Synchronization of Cell Populations in G1/S and G2/M Phases of the Cell Cycle
- The Myc World Within Reach
- 活细胞培养观察方法