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In nucleoli, the steady state of nucleolar proteins is leptomycin B‐sensitive
Author(s) -
Muro Eleonora,
Hoang Thang Q.,
JobartMalfait Aude,
HernandezVerdun Danièle
Publication year - 2008
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1042/bc20070117
Subject(s) - nucleolus , fibrillarin , nucleoplasm , biology , green fluorescent protein , microbiology and biotechnology , cajal body , nucleus , gene , biochemistry , rna , rna splicing
Background information . The nucleolus is a dynamic structure. It has been demonstrated that nucleolar proteins rapidly associate with and dissociate from nucleolar components in continuous exchanges with the nucleoplasm using GFP (green fluorescent protein)‐tagged proteins. However, how the exchanges within one nucleolus and between nucleoli within the nuclear volume occurred is still poorly understood. Results . The movement of PAGFP (photoactivatable GFP)‐tagged proteins that become visible after photoactivation can be followed. In the present study, we establish the protocol allowing quantification of the traffic of PAGFP‐tagged nucleolar proteins in nuclei containing two nucleoli. The traffic in the activated area, at the periphery of the activated area and to the neighbouring nucleolus is measured. Protein B23 is rapidly replaced in the activated area, and at the periphery of the activated area the steady state suggests intranucleolar recycling of B23; this recycling is LMB (leptomycin B)‐sensitive. The pool of activated B23 is equally distributed in the volume of the two nucleoli within 2 min. The three‐dimensional distribution of the proteins Nop52 and fibrillarin is less rapid than that of B23 but is also LMB‐sensitive. In contrast, traffic of fibrillarin from the nucleoli to the CB (Cajal body) was not modified by LMB. Conclusions . We propose that the steady state of nucleolar proteins in nucleoli depends on the affinity of the proteins for their partners and on intranucleolar recycling. This steady state can be impaired by LMB but not the uptake in the neighbouring nucleolus or the CB.