ELOVL1 production of C24 acyl-CoAs is linked to C24 sphingolipid synthesis

Ohno, Yusuke; Suto, Satoshi; Yamanaka, Masao; Mizutani, Yukiko; Mitsutake, Susumu; Igarashi, Yasuyuki; Sassa, Takayuki; Kihara, Akio

doi:10.1073/pnas.1005572107

Cited by 306 publications

(390 citation statements)

References 34 publications

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“…The codons under diversifying selection can be observed in Figure 4 and in Tables S4 and S5. Branch 4, which includes sequences that clustered with functionally characterized Elovl4 proteins (Ohno et al., 2010), was observed to have 15 codons under episodic diversifying selection with a posterior probability ≥ 0.95. The remaining branches are observed to have between 11 and 14 codons under episodic diversifying selection with a posterior probability ≥ 0.95.…”

Section: Resultsmentioning

confidence: 99%

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Surm

Toledo

Prentis

et al. 2018

Ecology and Evolution

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The biosynthesis of long‐chain polyunsaturated fatty acids (LC‐PUFAs, ≥ C20) is reliant on the action of desaturase and elongase enzymes, which are encoded by the fatty acyl desaturase (Fad) and elongation of very long‐chain fatty acid (Elovl) gene families, respectively. In Metazoa, research investigating the distribution and evolution of these gene families has been restricted largely to Bilateria. Here, we provide insights into the phylogenetic and molecular evolutionary histories of the Fad and Elovl gene families in Cnidaria, the sister phylum to Bilateria. Four model cnidarian genomes and six actiniarian transcriptomes were interrogated. Analysis of the fatty acid composition of a candidate cnidarian species, Actinia tenebrosa, was performed to determine the baseline profile of this species. Phylogenetic analysis revealed lineage‐specific gene duplication in actiniarians for both the Fad and Elovl gene families. Two distinct cnidarian Fad clades clustered with functionally characterized Δ5 and Δ6 proteins from fungal and plant species, respectively. Alternatively, only a single cnidarian Elovl clade clustered with functionally characterized Elovl proteins (Elovl4), while two additional clades were identified, one actiniarian‐specific (Novel ElovlA) and the another cnidarian‐specific (Novel ElovlB). In actiniarians, selection analyses revealed pervasive purifying selection acting on both gene families. However, codons in the Elovl gene family show patterns of nucleotide variation consistent with the action of episodic diversifying selection following gene duplication events. Significantly, these codons may encode amino acid residues that are functionally important for Elovl proteins to target and elongate different precursor fatty acids. In A. tenebrosa, the fatty acid analysis revealed an absence of LC‐PUFAs > C20 molecules and implies that the Elovl enzymes are not actively contributing to the elongation of these LC‐PUFAs. Overall, this study has revealed that actiniarians possess Fad and Elovl genes required for the biosynthesis of some LC‐PUFAs, and that these genes appear to be distinct from bilaterians.

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Section: Resultsmentioning

confidence: 99%

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Surm

Toledo

Prentis

et al. 2018

Ecology and Evolution

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“…Mammals have six ceramide synthases (CERS1-6) and seven FA elongases (ELOVL1-7), and each exhibits characteristic substrate specificity (Fig. S2B) (2,4,5,28). When CYP4F22 was expressed in these cells producing different ceramide species, ω-hydroxyceramides were produced in cells producing C26 ceramide (ELOVL1/CERS3 combination) and ≥C26 ceramides (the ELOVL4/CERS3 combination) but not in cells producing C22-C24 ceramides (the ELOVL1/CERS2 combination) (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…To examine this possibility, we prepared HEK 293T cells producing different sets of ceramides with specific chain lengths by introducing particular combinations of ceramide synthase and FA elongase (ELOVL1 and CERS2, C22-C24 ceramides; ELOVL1 and CERS3, C26 ceramide; and ELOVL4 and CERS3, ≥C26 ceramides) (Fig. 5C) (28,29). Mammals have six ceramide synthases (CERS1-6) and seven FA elongases (ELOVL1-7), and each exhibits characteristic substrate specificity (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Cells were labeled with 2 μCi [3-3 H]sphingosine (20 Ci/mmol; PerkinElmer Life Sciences) for 4 h at 37°C. Lipids were extracted as described previously (28,33) and separated by normal-phase TLC and reverse-phase TLC. Lipid Analysis Using LC-MS. FAs and ceramides prepared from cultured cells were analyzed by reversed-phase LC/MS using ultra-performance liquid chromatography (UPLC) coupled with electrospray ionization (ESI) tandem triple quadrupole MS (Xevo TQ-S; Waters).…”

Section: Methodsmentioning

confidence: 99%

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Essential role of the cytochrome P450 CYP4F22 in the production of acylceramide, the key lipid for skin permeability barrier formation

Ohno

Nakamichi

Ohkuni

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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A skin permeability barrier is essential for terrestrial animals, and its impairment causes several cutaneous disorders such as ichthyosis and atopic dermatitis. Although acylceramide is an important lipid for the skin permeability barrier, details of its production have yet to be determined, leaving the molecular mechanism of skin permeability barrier formation unclear. Here we identified the cytochrome P450 gene CYP4F22 (cytochrome P450, family 4, subfamily F, polypeptide 22) as the long-sought fatty acid ω-hydroxylase gene required for acylceramide production. CYP4F22 has been identified as one of the autosomal recessive congenital ichthyosis-causative genes. Ichthyosis-mutant proteins exhibited reduced enzyme activity, indicating correlation between activity and pathology. Furthermore, lipid analysis of a patient with ichthyosis showed a drastic decrease in acylceramide production. We determined that CYP4F22 was a type I membrane protein that locates in the endoplasmic reticulum (ER), suggesting that the ω-hydroxylation occurs on the cytoplasmic side of the ER. The preferred substrate of the CYP4F22 was fatty acids with a carbon chain length of 28 or more (≥C28). In conclusion, our findings demonstrate that CYP4F22 is an ultra-long-chain fatty acid ω-hydroxylase responsible for acylceramide production and provide important insights into the molecular mechanisms of skin permeability barrier formation. Furthermore, based on the results obtained here, we proposed a detailed reaction series for acylceramide production.acylceramide | ceramide | lipid | skin | sphingolipid

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“…Each amplified DNA fragment was cloned into the pGEM-T Easy vector (Promega, Fitchburg, WI). Human CERS2, CERS3, and CERS4 cDNAs cloned into the pGEM-T Easy vector have been described previously (29,30 Table 1) were constructed using the overlap extension PCR (for mutant (MT) forms of human CERS2-6) or QuikChange site-directed mutagenesis kit (Agilent Technologies, Santa Clara, CA) (for MTs of mouse Cers2), with appropriate WT plasmid as a template and primers (for CERS2 (4A), CERS2-4A-F and CERS2-4A-R; for CERS3 (S340A), CERS3-S340A-F and CERS3-S340A-R; for CERS4 (4A), CERS4 -4A-F and CERS4 -4A-R; for CERS5 (4A), CERS5-4A-F and CERS5-4A-R; for CERS6 (4A), CERS6 -4A-F and CERS6 -4A-R; for Cers2 (S341A), Cers2-S341-F and Cers2-S341-R; for Cers2 (T346A), Cers2-T346-F and Cers2-T346-R; for Cers2 (S348A), Cers2-S348-F and Cers2-S348-R; for Cers2 (S349A), Cers2-S349-F and Cers2-S349-R; for Cers2 (4A), Cers2-4A-F and Cers2-4A-R) ( Table 2). Each cDNA fragment was excised from the corresponding pGEM-T Easy-based plasmid and cloned into a mammalian expression vector pCE-puro HA-1, which is designed to produce a protein fused to an N-terminal HA tag under control of the human elongation factor 1␣ promoter (31).…”

Section: Methodsmentioning

confidence: 99%

Enzyme Activities of the Ceramide Synthases CERS2–6 Are Regulated by Phosphorylation in the C-terminal Region

Sassa

Hirayama

Kihara

2016

Journal of Biological Chemistry

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Ceramide and complex sphingolipids regulate important cellular functions including cell growth, apoptosis, and signaling. Dysregulation of sphingolipid metabolism leads to pathological consequences such as sphingolipidoses and insulin resistance. Ceramides in mammals vary greatly in their acyl-chain composition: six different ceramide synthase isozymes (CERS1-6) that exhibit distinct substrate specificity and tissue distribution account for this diversity. In the present study, we demonstrated that CERS2-6 were phosphorylated at the cytoplasmic C-terminal regions. Most of the phosphorylated residues conformed to a consensus motif for phosphorylation by casein kinase 2 (CK2), and treatment of cells with the CK2-specific inhibitor CX-4945 lowered the phosphorylation levels of CERS2, -4, -5, and -6. Phosphorylation of CERS2 was especially important for its catalytic activity, acting mainly by increasing its V max value. Phosphorylation modestly increased the catalytic activities of CERS4 and -5 and mildly increased those of CERS3 and -6. Dephosphorylation of endogenous ceramide synthases in the mouse brain led to severely reduced activity toward the Cers2 substrates C22:0/C24:0-CoAs and modestly reduced activity toward the Cers5/6 substrate C16:0-CoA. These results suggest that the phosphorylation of ceramide synthases may be a key regulatory point in the control of the distribution and levels of sphingolipids of various acyl-chain lengths.Sphingolipids, one of the major lipid constituents of eukaryotic membranes, mediate a number of cellular and physiological functions such as cell growth, apoptosis, immune responses, and the epidermal permeability barrier, whereas their abnormal metabolism is involved in diseases such as sphingolipidoses, neural disorders, and diabetes (1-7). Ceramides are located in the hub of sphingolipid metabolism. Ceramides are precursors for complex sphingolipids, whereas their hydrolysis releases a sphingoid long-chain base and a fatty acid (FA).2 The released long-chain base can then be recycled to synthesize new ceramide, or metabolized further into other lipids such as sphingosine 1-phosphate (8 -10). Ceramide synthases catalyze the formation of an amide bond between the long-chain base and the FA in the endoplasmic reticulum. In most tissues, the chain lengths of the FA moieties of ceramides are C16 -

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ELOVL1 production of C24 acyl-CoAs is linked to C24 sphingolipid synthesis

Cited by 306 publications

References 34 publications

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Essential role of the cytochrome P450 CYP4F22 in the production of acylceramide, the key lipid for skin permeability barrier formation

Enzyme Activities of the Ceramide Synthases CERS2–6 Are Regulated by Phosphorylation in the C-terminal Region

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