Tracking the interactions of rRNA processing proteins during nucleolar assembly in living cells

Reorganization of the nuclear machinery after mitosis is a fundamental but poorly understood process. Here, we investigate the recruitment of the nucleolar processing proteins in the nucleolus of living cells at the time of nucleus formation. We question the role of the prenucleolar bodies (PNBs), d...

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Published inMolecular biology of the cell Vol. 16; no. 6; pp. 2862 - 2871
Main Authors Angelier, Nicole, Tramier, Marc, Louvet, Emilie, Coppey-Moisan, Maïté, Savino, Tula M, De Mey, Jan R, Hernandez-Verdun, Danièle
Format Journal Article
LanguageEnglish
Published United States American Society for Cell Biology 01.06.2005
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Abstract Reorganization of the nuclear machinery after mitosis is a fundamental but poorly understood process. Here, we investigate the recruitment of the nucleolar processing proteins in the nucleolus of living cells at the time of nucleus formation. We question the role of the prenucleolar bodies (PNBs), during migration of the processing proteins from the chromosome periphery to sites of rDNA transcription. Surprisingly, early and late processing proteins pass through the same PNBs as demonstrated by rapid two-color four-dimensional imaging and quantification, whereas a different order of processing protein recruitment into nucleoli is supported by differential sorting. Protein interactions along the recruitment pathway were investigated using a promising time-lapse analysis of fluorescence resonance energy transfer. For the first time, it was possible to detect in living cells the interactions between proteins of the same rRNA processing machinery in nucleoli. Interestingly interactions between such proteins also occur in PNBs but not at the chromosome periphery. The dynamics of these interactions suggests that PNBs are preassembly platforms for rRNA processing complexes.
AbstractList Reorganization of the nuclear machinery after mitosis is a fundamental but poorly understood process. Here, we investigate the recruitment of the nucleolar processing proteins in the nucleolus of living cells at the time of nucleus formation. We question the role of the prenucleolar bodies (PNBs), during migration of the processing proteins from the chromosome periphery to sites of rDNA transcription. Surprisingly, early and late processing proteins pass through the same PNBs as demonstrated by rapid two-color four-dimensional imaging and quantification, whereas a different order of processing protein recruitment into nucleoli is supported by differential sorting. Protein interactions along the recruitment pathway were investigated using a promising time-lapse analysis of fluorescence resonance energy transfer. For the first time, it was possible to detect in living cells the interactions between proteins of the same rRNA processing machinery in nucleoli. Interestingly interactions between such proteins also occur in PNBs but not at the chromosome periphery. The dynamics of these interactions suggests that PNBs are preassembly platforms for rRNA processing complexes.
Author Hernandez-Verdun, Danièle
Tramier, Marc
Savino, Tula M
Angelier, Nicole
Louvet, Emilie
Coppey-Moisan, Maïté
De Mey, Jan R
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  givenname: Nicole
  surname: Angelier
  fullname: Angelier, Nicole
  organization: Nuclei and Cell Cycle Laboratory, Institut Jacques Monod, Centre National de la Recherche Scientifique, University Paris VI and Paris VII, 75251 Paris, France
– sequence: 2
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Snippet Reorganization of the nuclear machinery after mitosis is a fundamental but poorly understood process. Here, we investigate the recruitment of the nucleolar...
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SubjectTerms Anaphase
Cell Line, Transformed
Cell Nucleolus - metabolism
Cellular Biology
Chromosomal Proteins, Non-Histone - metabolism
Fluorescence Resonance Energy Transfer
G1 Phase
Green Fluorescent Proteins - metabolism
HeLa Cells
Humans
Image Processing, Computer-Assisted
Kinetics
Life Sciences
Microscopy, Video
Mitosis
Models, Biological
Nuclear Proteins - metabolism
Protein Processing, Post-Translational
RNA, Ribosomal - metabolism
Telophase
Transcription, Genetic
Title Tracking the interactions of rRNA processing proteins during nucleolar assembly in living cells
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