title: The rate-limiting enzyme in phosphatidylcholine synthesis is associated with nuclear speckles under stress conditions.
abstract: Phosphatidylcholine (PtdCho) is the most abundant phospholipid in eukaryotic membranes and its biosynthetic pathway is generally controlled by CTP:Phosphocholine Cytidylyltransferase (CCT), which is considered the rate-limiting enzyme. CCT is an amphitropic protein, whose enzymatic activity is commonly associated with endoplasmic reticulum (ER) translocation; however, most of the enzyme is intranuclearly located. Here we demonstrate that CCTα is concentrated in the nucleoplasm of MDCK cells. Confocal immunofluorescence revealed that extracellular hypertonicity shifted the diffuse intranuclear distribution of the enzyme to intranuclear domains in a foci pattern. One population of CCTα foci colocalised and interacted with lamin A/C speckles, which also contained the pre-mRNA processing factor SC-35, and was resistant to detergent and salt extraction. The lamin A/C silencing allowed us to visualise a second more labile population of CCTα foci that consisted of lamin A/C-independent foci non-resistant to extraction. We demonstrated that CCTα translocation is not restricted to its redistribution from the nucleus to the ER and that intranuclear redistribution must thus be considered. We suggest that the intranuclear organelle distribution of CCTα is a novel mechanism for the regulation of enzyme activity.
authors.affiliation: Department of Biological Sciences, University of Biological Sciences, University of Buenos Aires, Buenos Aires, Argentina.
authors.forename: Nicolás O
authors.initials: NO
authors.lastname: Favale
authors.forename: María C
authors.initials: MC
authors.lastname: Fernández-Tome
authors.forename: Lucila G
authors.initials: LG
authors.lastname: Pescio
authors.forename: Norma B
authors.initials: NB
authors.lastname: Sterin-Speziale
mesh_terms: D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D019916:Choline-Phosphate Cytidylyltransferase / Q000737:chemistry / Q000502:physiology*; D003593:Cytoplasm / Q000378:metabolism; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004798:Enzymes / Q000737:chemistry*; D020868:Gene Silencing; D034904:Lamin Type A / Q000737:chemistry; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D021381:Protein Transport; D013997:Time Factors
chemical_list: D004798:Enzymes; D034904:Lamin Type A; D010713:Phosphatidylcholines; C440308:lamin C; D019916:Choline-Phosphate Cytidylyltransferase