Spatial link between nucleoli and expression of the Zac1 gene.
Eukaryotic genomes are highly organized within the cell nucleus. Genome organization not only implies the preferential positioning of genetic elements in the interphase nucleus but also the topographic distribution of biological processes. We have investigated the relationship between spatial organization and genome function in single cells. Myc, c-Met, Igf2r, Asb4, and Zac1 genes have the same radial distribution, but they are not positioned in close proximity with respect to each other. Three-dimensional mapping of their transcription sites uncovered a gene-specific pattern of relative positioning with respect to the nucleolus. We found that the Zac1 gene transcription preferentially occurs juxtaposed to the nucleolus, and that its mRNA accumulates at this site of transcription. Nucleoli isolation followed by qRT-PCR provided evidence for a physical interaction between Zac1 mRNA and the nucleolus. Actinomycin-D treatment induced disassembly of the nucleolus, loss of the RNA-FISH signal, and dramatic increase of the ZAC protein level. However, inhibition of RNA polymerase II had no effect over the Zac1 FISH signal and the protein expression. Induction of cell cycle arrest, which involves participation of the ZAC protein, provoked mRNA release from its retention site and protein synthesis. Our data demonstrate that Zac1 mRNA preferentially accumulates in close proximity to nucleoli within the cell nucleus. In addition, our results suggest a functional link between such spatial distribution and protein expression.
Royo
Félix
F
Cytogenomics, CIC bioGUNE-CIBEREHD, Par. Tec. Bizkaia Ed. 801 A, Derio, Spain.
Paz
Nerea
N
Espinosa
Luis
L
McQueen
Philip G
PG
Vellón
Luciano
L
Parada
Luis A
LA
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D002466:Cell Nucleolus / Q000187:drug effects / Q000235:genetics*; D045584:Chromosome Positioning / Q000187:drug effects; D033481:Chromosomes, Mammalian / Q000235:genetics; D004249:DNA Damage; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D016147:Genes, Tumor Suppressor; D056426:Genetic Loci / Q000235:genetics; D051379:Mice; D014176:Protein Biosynthesis / Q000187:drug effects; D012318:RNA Polymerase I / Q000037:antagonists & inhibitors; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D018797:Cell Cycle Proteins; D004791:Enzyme Inhibitors; C498423:Plagl1 protein, mouse; D012333:RNA, Messenger; D014157:Transcription Factors; D012319:RNA Polymerase II; D012318:RNA Polymerase I