Abstract

Nucleogenesis is the cellular event responsible for the formation of the new nucleoli at the end of mitosis. This process depends on the synthesis and processing of ribosomal RNA (rRNA) and, in some eukaryotes, the transfer of nucleolar material contained in prenucleolar bodies (PNBs) to active transcription sites. The lack of a comprehensive description of the nucleolus throughout the cell cycle of the human pathogen Leishmania major prompted us to analyze the distribution of nucleolar protein 56 (Nop56) during interphase and mitosis in the promastigote stage of the parasite. By in silico analysis we show that the orthologue of Nop56 in L. major (LmNop56) contains the three characteristic Nop56 domains and that its predicted three-dimensional structure is also conserved. Fluorescence microscopy observations indicate that the nucleolar localization of LmNop56 is similar, but not identical, to that of the nucleolar protein Elp3b. Notably, unlike other nucleolar proteins, LmNop56 remains associated with the nucleolus in nonproliferative cells. Moreover, epifluorescent images indicate the preservation of the nucleolar structure throughout the closed nuclear division. Experiments performed with the related parasite Trypanosoma brucei show that nucleolar division is carried out by an analogous mechanism.

Highlights

  • The cell nucleus contains a collection of nonmembranebound nuclear bodies (NBs) that participate in the regulation of essential functions, such as gene expression [1, 2]

  • The nucleolar cycle begins during the early stages of nuclear division, when several key nucleolar proteins involved in rDNA transcription and ribosomal RNA (rRNA) processing are negatively modulated by specific phosphorylation carried out by the cyclin B-dependent kinase 1 pathway [9,10,11]

  • Multiple sequence alignments indicated that LmNop56 is ∼80% identical to the T. brucei and T. cruzi orthologues, and 46 and 48% identical to nucleolar protein 56 (Nop56) from human and yeast, respectively

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Summary

Introduction

The cell nucleus contains a collection of nonmembranebound nuclear bodies (NBs) that participate in the regulation of essential functions, such as gene expression [1, 2]. The nucleolar cycle begins during the early stages of nuclear division, when several key nucleolar proteins involved in rDNA transcription and rRNA processing are negatively modulated by specific phosphorylation carried out by the cyclin B-dependent kinase 1 pathway [9,10,11]. While proteins that participate in rDNA transcription remain attached to nucleolar organizer regions (NORs), rRNA processing proteins and small nucleolar RNAs (snoRNAs) as well as preserved prerRNAs localize to the cytoplasm and progressively accumulate along the entire periphery of condensed chromosomes, forming part of the perichromosomal compartment (PC) [12,13,14,15]. The nucleolar material accumulates in intermediate nuclear structures called prenucleolar bodies (PNBs), before being released into transcriptionally active NORs, which are chromosomal loci where

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