Protein NO52--a constitutive nucleolar component sharing high sequence homologies to protein NO66.
The nucleolus is the most prominent intranuclear structure of almost all protein-synthesizing cells. It compromises a well-defined functional compartmentalization and a high complexity of molecular constituents. Here, we report on the identification and molecular characterization of a novel constitutive nucleolar component--protein NO52--that is present in diverse species from Xenopus laevis to human. The cDNA-deduced amino acid sequence of protein NO52 defines a polypeptide of a calculated mass of 52.8 kDa and an isoelectric point of 6.7. Inspection of the primary sequence disclosed that the protein contains a JmjC domain and is highly sequence-related to the recently described nucleolar protein NO66. Immunolocalization studies revealed that protein NO52 is highly concentrated in the granular component of nucleoli and this characteristic intranuclear distribution is significantly affected by treatment of cells with (i) RNase A, (ii) actinomycin D and (iii) serum starvation. Interestingly, protein NO52 has been identified as a constituent of free preribosomal particles but is absent from cytoplasmic ribosomes. Analyses of immunocomplexes isolated from cellular extracts with an NO52-specific antibody by MALDI mass spectrometry further confirmed the interaction of protein NO52 with various ribosomal proteins as well as with a distinct set of non-ribosomal nucleolar proteins. The dependence of the nucleolar accumulation of the protein on ongoing rRNA transcription and the cellular metabolic state strongly suggest that protein NO52 is directly involved in ribosome biogenesis, most likely during the assembly process of preribosomal particles.
Eilbracht
Jens
J
Division of Cell Biology, German Cancer Research Center, Heidelberg, Germany.
Kneissel
Sandra
S
Hofmann
Astrid
A
Schmidt-Zachmann
Marion S
MS
D000595:Amino Acid Sequence; D000818:Animals; D002466:Cell Nucleolus / Q000502:physiology*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics* / Q000502:physiology; D049308:Dioxygenases; D056466:Histone Demethylases; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data
D002868:Chromosomal Proteins, Non-Histone; D049308:Dioxygenases; D056466:Histone Demethylases; C492859:RIOX1 protein, human