CGI-58 Protein Acts as a Positive Regulator of Triacylglycerol Accumulation in Phaeodactylum tricornutum
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Comparative gene identification-58 (CGI-58) is an activating protein of triacylglycerol (TAG) lipase. It has a variety of catalytic activities whereby it may play different roles in diverse organisms. In this study, a homolog of CGI-58 in Phaeodactylum tricornutum (PtCGI-58) was identified. PtCGI-58 was localized in mitochondria by GFP fusion protein analysis, which is different from the reported subcellular localization of CGI-58 in animals and plants. Respectively, PtCGI-58 overexpression resulted in increased neutral lipid content and TAG accumulation by 42-46% and 21-32%. Likewise, it also increased the relative content of eicosapentaenoic acid (EPA), and in particular, the EPA content in TAGs almost doubled. Transcript levels of genes involved in de novo fatty acid synthesis and mitochondrial β-oxidation were significantly upregulated in PtCGI-58 overexpression strains compared with wild-type cells. Our findings suggest that PtCGI-58 may mediate the breakdown of lipids in mitochondria and the recycling of acyl chains derived from mitochondrial β-oxidation into TAG biosynthesis. Moreover, this study potentially illuminates new functions for CGI-58 in lipid homeostasis and provides a strategy to enrich EPA in algal TAGs.
Abstract
Comparative gene identification-58 (CGI-58) is an activating protein of triacylglycerol (TAG) lipase. It has a variety of catalytic activities whereby it may play different roles in diverse organisms. In this study, a homolog of CGI-58 in Phaeodactylum tricornutum (PtCGI-58) was identified. PtCGI-58 was localized in mitochondria by GFP fusion protein analysis, which is different from the reported subcellular localization of CGI-58 in animals and plants. Respectively, PtCGI-58 overexpression resulted in increased neutral lipid content and TAG accumulation by 42-46% and 21-32%. Likewise, it also increased the relative content of eicosapentaenoic acid (EPA), and in particular, the EPA content in TAGs almost doubled. Transcript levels of genes involved in de novo fatty acid synthesis and mitochondrial β-oxidation were significantly upregulated in PtCGI-58 overexpression strains compared with wild-type cells. Our findings suggest that PtCGI-58 may mediate the breakdown of lipids in mitochondria and the recycling of acyl chains derived from mitochondrial β-oxidation into TAG biosynthesis. Moreover, this study potentially illuminates new functions for CGI-58 in lipid homeostasis and provides a strategy to enrich EPA in algal TAGs.
참고문헌 (42)
- Field CB, "Primary production of the biosphere: integrating terrestrial and oceanic components", Science, 281, pp.237~240 (1998)
- Falkowski PG, "The evolution of modern eukaryotic phytoplankton", Science, 305, pp.354~360 (2004)
- Du Z-Y, "Lipids in Plant and Algae Development", pp.179~205 (2016), Springer
- Yu E, "Triacylglycerol accumulation and profiling in the model diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum (Baccilariophyceae) during starvation", J. Appl. Phycol, 21, pp.669~681 (2009)
- Kong F, "Lipid catabolism in microalgae", New Phytol, 218, pp.1340~1348 (2018)
- Casas-Godoy L, "Lipases and Phospholipases", pp.3~30 (2012), Humana Press
- Oberer M, "Recent insights into the structure and function of CGI-58", Curr. Opin. Lipidol, 22, pp.149~158 (2011)
- Yu L, "CGI-58: versatile regulator of intracellular lipid droplet homeostasis", Adv. Exp. Med. Biol, 1276, pp.197~222 (2020)
- Yamaguchi T, "CGI-58 interacts with Perilipin and is localized to lipid droplets: possible involvement of CGI-58 mislocalization in Chanarin-Dorfman syndrome", J. Biol. Chem, 279, pp.30490~30497 (2004)
- Subramanian V, "Perilipin A mediates the reversible binding of CGI-58 to lipid droplets in 3T3-L1 adipocytes", J. Biol. Chem, 279, pp.42062~42071 (2004)
- Radner FPW, "Growth retardation, impaired triacylglycerol catabolism, hepatic steatosis, and lethal skin barrier defect in mice lacking comparative gene identification-58 (CGI-58)", J. Biol. Chem, 285, pp.7300~7311 (2010)
- James CN, "Disruption of the Arabidopsis CGI-58 homologue produces Chanarin-Dorfman-like lipid droplet accumulation in plants", Proc. Nat. Acad. Sci. USA, 107, pp.17833~17838 (2010)
- Park S, "The α", β hydrolase CGI-58 and peroxisomal transport protein PXA1 coregulate lipid homeostasis and signaling in Arabidopsis, 2013, pp.25, 1726~1739
- Trentacoste EM, "Metabolic engineering of lipid catabolism increases microalgal lipid accumulation without compromising growth", Proc. Nat. Acad. Sci. USA, 110, pp.19748~19753 (2013)
- Ghosh AK, "YLR099C (ICT1) encodes a soluble Acyl-CoA-dependent lysophosphatidic acid acyltransferase responsible for enhanced phospholipid synthesis on organic solvent stress in Saccharomyces cerevisiae", J. Biol. Chem, 283, pp.9768~9775 (2008)
- Montero-Moran G, "CGI-58", ABHD5 is a coenzyme Adependent lysophosphatidic acid acyltransferase, 2010, pp.51, 709~719
- Ghosh AK, "At4g24160, a soluble acyl-coenzyme A-dependent lysophosphatidic acid acyltransferase", Plant Physiol, 151, pp.869~881 (2009)
- McMahon D, "Comparative gene identification 58", α, J. Lipid Res, pp.2014, 55[DOI]
- Guillard RRL, "Culture of Marine Invertebrate Animals", pp.29~60 (1975), Plenum Press
- Collos Y, "An optical method for the rapid measurement of micromolar concentrations of nitrate in marine phytoplankton cultures", J. Appl. Phycol, 11, pp.179~184 (1999)
- Maheswari U, "Digital expression profiling of novel diatom transcripts provides insight into their biological functions", Genome Biol, 11, pp.R85~ (2010)
- Guindon S, "PHYML Online—a web server for fast maximum likelihood-based phylogenetic inference", Nucleic Acids Res, 33, pp.W557~W559 (2005)
- Zaslavskaia LA, "Transformation of the diatom Phaeodactylum tricornutum (Bacillariophyceae) with a variety of selectable marker and reporter genes", J. Phycol, 36, pp.379~386 (2000)
- Zhang C, "High-efficiency nuclear transformation of the diatom Phaeodactylum tricornutum by electroporation", Mar. Genomics, 16, pp.63~66 (2014)
- Siaut M, "Molecular toolbox for studying diatom biology in Phaeodactylum tricornutum", Gene, 406, pp.23~35 (2007)
- Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method", Methods, 25, pp.402~408 (2001)
- Bligh EG, "A rapid method of lipid extraction and purification", Can. J. Biochem. Physiol, 37, pp.911~917 (1959)
- Reiser S, "Isolation of mutants of Acinetobacter calcoaceticus deficient in wax ester synthesis and complementation of one mutation with a gene encoding a fatty acyl-coenzyme a reductase", J. Bacteriol, 179, pp.2969~2975 (1997)
- Xie Y, "Ultrasound-assisted one-phase solvent extraction coupled with liquid chromatography-quadrupole time-of-flight mass spectrometry for efficient profiling of egg yolk lipids", Food Chem, 319, pp.126547~ (2020)
- Gruber A, "The N-terminal region of comparative gene identification-58 (CGI-58) is important for lipid droplet binding and activation of adipose triglyceride lipase", J. Biol. Chem, 285, pp.12289~12298 (2010)
- Schrag JD, "Lipases and αβ hydrolase fold", Meth. Enzymol, 284, pp.85~107 (1997)
- Ghosh AK, "CGI-58, the causative gene for Chanarin-Dorfman syndrome, mediates acylation of lysophosphatidic acid", J. Biol. Chem, 283, pp.24525~24533 (2008)
- Kurat CF, "Obese yeast: triglyceride lipolysis is functionally conserved from mammals to yeast", J. Biol. Chem, 281, pp.491~500 (2006)
- Li X, "Identification of the triacylglycerol lipase in the chloroplast envelope of the diatom Phaeodactylum tricornutum", Algal Res, 33, pp.440~447 (2018)
- Barka F, "Identification of a triacylglycerol lipase in the diatom Phaeodactylum tricornutum", Biochim. Biophys. Acta Mol. Cell Biol. Lipids, 1861, pp.239~248 (2016)
- Jallet D, "Mitochondrial fatty acid β-oxidation is required for storage-lipid catabolism in a marine diatom", New Phytol, 228, pp.946~958 (2020)
- Hao X, "Multiplex CRISPR", Cas9 editing of the long-chain acyl-CoA synthetase family in the diatom Phaeodactylum tricornutum reveals that mitochondrial ptACSL3 is involved in the synthesis of storage lipids, 2022, pp.233, 1797~1812
- Bates PD, "Analysis of acyl fluxes through multiple pathways of triacylglycerol synthesis in developing soybean embryos", Plant Physiol, 150, pp.55~72 (2009)
- Bates PD, "Acyl editing and headgroup exchange are the major mechanisms that direct polyunsaturated fatty acid flux into triacylglycerols", Plant Physiol, 160, pp.1530~1539 (2012)
- Khatib A, "Reassessing the potential activities of plant CGI-58 protein", PLoS One, 11, pp.e0145806~ (2016)
- Abida H, "Membrane glycerolipid remodeling triggered by nitrogen and phosphorus starvation in Phaeodactylum tricornutum", Plant Physiol, 167, pp.118~136 (2015)
- Zhang J, "Comparative gene identification-58 (CGI-58) promotes autophagy as a putative lysophosphatidylglycerol acyltransferase", J. Biol. Chem, 289, pp.33044~33053 (2014)
