CTP synthase is an essential enzyme that plays a key role in energy metabolism. Several independent studies have demonstrated that CTP synthase can form an evolutionarily conserved subcellular structure termed cytoophidium. In budding yeast, there are two isoforms of CTP synthase and both isoforms localize in cytoophidium. However, little is known about the distribution of CTP synthase isoforms in Drosophila melanogaster . Here, we report that three transcripts generated at the CTP synthase gene locus exhibit different expression profiles, and three isoforms encoded by this gene locus show a distinct subcellular distribution. While isoform A localizes in the nucleus, isoform B distributes diffusely in the cytoplasm, and only isoform C forms the cytoophidium. In the two isoform C-specific mutants, cytoophidia disappear in the germline cells. Although isoform A does not localize to the cytoophidium, a mutation disrupting mostly isoform A expression results in the disassembly of cytoophidia. Overexpression of isoform C can induce the growth of the cytoophidium in a cell-autonomous manner. Ectopic expression of the cytoophidium-forming isoform does not cause any defect in the embryos. In addition, we identify that a small segment at the amino terminus of isoform C is necessary but not sufficient for cytoophidium formation. Finally, we demonstrate that an excess of the synthetase domain of CTP synthase disrupts cytoophidium formation. Thus, the study of multiple isoforms of CTP synthase in Drosophila provides a good opportunity to dissect the biogenesis and function of the cytoophidum in a genetically tractable organism. Author Summary DNA and RNA are made up from basic building blocks called nucleotides. Those nucleotides also play essential roles in many other biological processes. To separate biological processes within a cell is an important feature of all cell types. For example, mitochondria are specialized structures that contain ATP synthase, the enzyme that makes the nucleotide ATP. While mitochondria and ATP synthase have been studied for about 100 years, it was only very recently that we realized that there are specialized subcellular structures that contain CTP synthase, the enzyme that makes up another basic nucleotide CTP. Several independent studies have shown that CTP synthase molecules can form a filamentous structure called the cytoophidium (meaning “cellular snake” in Greek) or CTP synthase filament in bacteria, budding yeasts, fruit flies, and rat and human cells. In budding yeast, there are two isoforms of CTP synthase and both isoforms localize in the cytoophidium. Here, we report that three CTP synthase isoforms in fruit flies show a distinct subcellular distribution and only one isoform forms the cytoophidium. Thus, the study of multiple isoforms of CTP synthase in the fruit fly gives us a good way to begin to learn how and why CTP synthase molecules form this snake-like structure.
【초록키워드】 ectopic expression, Structure, Mitochondria, Mutation, metabolism, cells, energy metabolism, mutants, Bacteria, ATP, distribution, expression, yeast, nucleotide, nucleotides, cell types, Drosophila melanogaster, cytoplasm, Drosophila, locus, biological processes, enzyme, growth, Author, domain, human cells, expression profiles, organism, overexpression, nucleus, embryos, ATP synthase, biogenesis, while, transcript, block, isoform, DNA and RNA, Cell, independent, subcellular distribution, shown, different, identify, conserved, example, addition, form, provide, demonstrated, induce, dissect, disrupt, disrupting, Synthase, biological processe, synthase gene, subcellular structure, 【제목키워드】 Drosophila melanogaster,