Generation of a ceramide synthase 6 mouse lacking the DDRSDIE C-terminal motif.

Autor: Kim J; Department Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel., Pewzner-Jung Y; Department Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel., Joseph T; Department Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel., Ben-Dor S; Life Sciences Core Facilities, Weizmann Institute of Science, Rehovot, Israel., Futerman AH; Department Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2022 Jul 18; Vol. 17 (7), pp. e0271675. Date of Electronic Publication: 2022 Jul 18 (Print Publication: 2022).
DOI: 10.1371/journal.pone.0271675
Abstrakt: The important membrane lipid, ceramide, is generated by a family of homologous enzymes, the ceramide synthases (CerSs), multi-spanning membrane proteins located in the endoplasmic reticulum. Six CerS isoforms exist in mammals with each using a subset of acyl-CoAs for (dihydro)ceramide synthesis. A number of mice have been generated in which one or other CerS has been genetically manipulated, including complete knock-outs, with each displaying phenotypes concomitant with the expression levels of the CerS in question and the presumed biological function of the ceramide species that it generates. We recently described a short C-terminal motif in the CerS which is involved in CerS dimer formation; deleting this motif had no effect on the ability of the CerS to synthesize ceramide in vitro. In the current study, we generated a CerS6 mouse using CRISPR-Cas9, in which the DDRSDIE motif was replaced by ADAAAIA. While levels of CerS6ADAAAIA expression were unaffected in the CerS6ADAAAIA mouse, and CerS6ADAAAIA was able to generate C16-ceramide in vitro, ceramide levels were significantly reduced in the CerS6ADAAAIA mouse, suggesting that replacing this motif affects an as-yet unknown mechanism of regulation of ceramide synthesis via the DDRSDIE motif in vivo. Crossing CerS6ADAAAIA mice with CerS5 null mice led to generation of viable mice in which C16-ceramide levels were reduced by up to 90%, suggesting that depletion of C16-ceramide levels is compensated for by other ceramide species with different acyl chain lengths.
Competing Interests: The authors declare that no competing interests exist.
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje