Visual Biology of Nuclear Dynamics: From Micro- to Nano-dynamics of Nuclear Components
When you look at an interphase nucleus in a living cell through a light microscope, you will see a round, static organelle separated from the cytoplasm. If you continue the live cell observation, you will easily learn that the cell nucleus does not undergo any significant morphological changes until it reaches the mitosis, where the nuclear envelope and the chromosomes show dynamic structural changes. Because of these morphological properties, the cell nucleus had previously been considered a “container” of genome that provides an enclosed space for genomic events to be carried out. However, recent progress in molecular and cellular biological approaches has led to the revelation that the cell nucleus is composed of various kinds of different “compartments,” each of which is supposed to have a distinct “structure” and “function.” These include promeyelocytic leukemia (PML) bodies, Cajal bodies, nucleolus, nuclear speckles, and nuclear foci (see figure in the Preface). Recent developments in various fluo-rescence observation techniques have revealed that these compartments are moving within a nucleus and there is a constant flow of proteins between nucleoplasm and these compartments (Fig. 1).In this chapter, therecent progress in various “visualization techniques” will be reviewed and how these techniques have been utilized to visualize the structures and the dynamics of the inner nuclear compartments and chromosomes will be described.
KeywordsAtomic Force Microscopy Fluorescent Resonance Energy Transfer Nuclear Pore Complex Fluorescence Recovery After Photobleaching Chromosome Territory
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- Allen MJ, Lee C, Lee JD, Pogany GC, Balooch M, Siekhaus WJ, Balhorn R (1993b) Atomic force microscopy of mammalian sperm chromatin. Chromo-soma 102:623–630Google Scholar
- Ancelin K, Brunori M, Bauwens S, Koering CE, Brun C, Ricoul M, Pommier JP, Sabatier L, Gilson E (2002) Targeting assay to study the cis functions of hu-man telomeric proteins: evidence for inhibition of telomerase by TRF1 and for activation of telomere degradation by TRF2. Mol Cell Biol 22:3474–3487PubMedGoogle Scholar
- Day RN, Periasamy A, Schaufele F (2001) Fluorescence resonance energy transfer microscopy of localized protein interactions in the living cell nucleus. Meth-ods 25:4–18Google Scholar
- Greene EC, Mizuuchi K (2004) Visualizing the assembly and disassembly mecha-nisms of the MuB transposition targeting complex. J Biol ChemGoogle Scholar
- Hizume K, Yoshimura SH, Wada H, Takeyasu K (2003) Atomic force microscopy demonstrates a critical role of DNA superhelicity in the nucleosome dynamics. Cell Biochem. Biophys 40:249–261Google Scholar
- Marcello A, Cinelli RA, Ferrari A, Signorelli A, Tyagi M, Pellegrini V, Beltram F, Giacca M (2001) Visualization of in vivo direct interaction between HIV-1 TAT and human cyclin Tl in specific subcellular compartments by fluores-cence resonance energy transfer. J Biol Chem 276:39220–39225PubMedGoogle Scholar
- Sleeman JE, Trinkle-Mulcahy L, Prescott AR, Ogg SC, Lamond AI (2003) Cajal body proteins SMN and Coilin show differential dynamic behaviour in vivo. J CellSci 116:2039–2050Google Scholar
- Winkler R, Perner B, Rapp A, Durm M, Cremer C, Greulich KO, Hausmann M (2003) Labeling quality and chromosome morphology after low temperature FISH analysed by scanning far-field and near-field optical microscopy. J Mi-crosc 209:23–33Google Scholar
- Yoshimura SH, Yoshida C, Igarashi K, Takeyasu K (2000b) Atomic force microscopy proposes a “kiss and pull” mechanism for enhancer function. J Electron Microsc (Tokyo) 49:407–413Google Scholar
- Yoshimura SH, Kim J, Takeyasu K (2003) On-substrate lysis treatment combined with scanning probe microscopy revealed chromosome structures in eukaryotes and prokaryotes. J Electron Microsc (Tokyo) 52:415–423Google Scholar
- Yoshino T, Sugiyama S, Hagiwara S, Ushiki T, Ohtani T (2002) Simultaneous collection of topographic and fluorescent images of barley chromosomes by scanning near-field optical/atomic force microscopy. J Electron Microsc (To-kyo) 51:199–203Google Scholar