Abstract
Phosphatidate phosphatase-1 (PAP1) enzymes (yeast Pah1p/Smp2p, mammalian lipin1-3) have a key role in lipid homeostasis by controlling the relative proportions of its substrate phosphatidate (PA) and its product diacylglycerol (DAG). Recent investigation shows that mammalian lipin-1 complements phenotypes exhibited by yeast pah1Δ mutant cells, which indicates the functions of PAP1 enzymes are evolutionarily conserved. The observation was confirmed after transformation of human LPIN1 into PAH1-defective yeast, which resulted in human LPIN1-induced accumulation of triacylglycerol (TAG) and lipid droplet formation. In double mutants lacking Tgl3p and Tgl4p, overexpression of PAH1 or LPIN1-induced TAG accumulation and excessive obesity. Furthermore, the obese yeast was used as a model to study the anti-obesity effects of PAP1 activity inhibitors, including propranolol and clenbuterol. The data showed that the inhibitors significantly suppressed TAG accumulation and lipid droplets formation. These findings demonstrate that LPIN1 plays a functional role in lipid synthesis and storage, a role which is highly conserved from human to yeast. Inhibition of TAG synthesis will become an efficacious treatment strategy for obesity and our excessive obesity model will provide a very useful tool for discovery of new anti-obesity drugs in the future.
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Abbreviations
- ATGL:
-
Adipose triglyceride lipase
- DAG:
-
Diacylglycerol
- DIC:
-
Differential interference contrast microscopy
- HSL:
-
Hormone-sensitive lipase
- PA:
-
Phosphatidate
- PAP:
-
Phosphatidate phosphatase
- TAG:
-
Triglyceride
- TLC:
-
Thin-layer chromatography
- WT:
-
Wild type
References
Adeyo, O., Horn, P. J., Lee, S. K., Binns, D. D., Chandrahas, A., Chapman, K. D., Goodman, J. M. (2011) The yeast lipin orthologue Pah1p is important for biogenesis of lipid droplets. J. Cell Biol. 192, 1043–1055.
Bilyk, A., Piazza, G., Bistline, R., Jr., Haas, M. (1991) Separation of cholesterol, and fatty acylglycerols, acids and amides by thin-layer chromatography. Lipids 26, 405–406.
Brachmann, C. B., Davies, A., Cost, G. J. Caputo, E., Li, J., Hieter, P., Boeke, J. D. (1998) Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14, 115–132.
Carman, G. M., Han, G. S. (2006) Roles of phosphatidate phosphatase enzymes in lipid metabolism. Trends Biochem. Sci. 31, 694–699.
Choi, H. S., Su, W. M., Han, G. S., Plote, D., Xu, Z., Carman, G. M. (2012) Pho85p-Pho80p phosphorylation of yeast Pah1p phosphatidate phosphatase regulates its activity, location, abundance, and function in lipid metabolism. J. Biol. Chem. 287, 11290–11301.
Csaki, L. S., Reue, K. (2010) Lipins: multifunctional lipid metabolism proteins. Ann. Rev. Nutr. 30, 257–272.
Gietz, R. D., Akio, S. (1988) New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene 74, 527–534.
Grimsey, N., Han, G. S., O’hara, L., Rochford, J. J., Carman, G. M., Siniossoglou, S. (2008) Temporal and spatial regulation of the phosphatidate phosphatases lipins 1 and 2. J. Biol. Chem. 283, 29166–29174.
Guo, Y., Cordes, K. R., Farese, R. V., Jr., Walther, T. C. (2009) Lipid droplets at a glance. J. Cell Sci. 122, 749–752.
Huang, Z., Sucgang, R. S., Lin, Y. Y., Shi, X., Boeke, J. D., Pan, X. (2008) Plasmid-chromosome shuffling for non-deletion alleles in yeast. Nat. Meth. 5, 167–169.
Kitazono, A. A. (2009) Improved gap-repair cloning method that uses oligonucleotides to target cognate sequences. Yeast 26, 497–505.
Kohlwein, S. D. (2010) Triacylglycerol homeostasis: insights from yeast. J. Biol. Chem. 285, 15663–15667.
Kurat, C. F., Natter, K., Petschnigg, J., Wolinski, H., Scheuringer, K., Scholz, H., Zimmermann, R., Leber, R., Zechner, R., Kohlwein, S. D. (2006) Obese yeast: triglyceride lipolysis is functionally conserved from mammals to yeast. J. Biol. Chem. 281, 491–500.
Nanjundan, M., Possmayer, F. (2003) Pulmonary phosphatidic acid phosphatase and lipid phosphate phosphohydrolase. Am. J. Physiol. Lung Cell. Mol. Physiol. 284, L1–L23.
O’hara, L., Han, G. S., Peak-Chew, S., Grimsey, N., Carman, G. M., Siniossoglou, S. (2006) Control of phospholipid synthesis by phosphorylation of the yeast lipin Pah1p/Smp2p Mg2+-dependent phosphatidate phosphatase. J. Biol. Chem. 281, 34537–34548.
Phan, J., Reue, K. (2005) Lipin, a lipodystrophy and obesity gene. Cell Metab. 1, 73–83.
Pyne, S., Long, J. S., Ktistakis, N. T., Pyne, N. J. (2005) Lipid phosphate phosphatases and lipid phosphate signalling. Biochem. Soc. Trans. 33, 1370–1374.
Reue, K., Dwyer, J. R. (2009) Lipin proteins and metabolic homeostasis. J. Lipid Res. 50 Suppl, S109–S114.
Santos-Rosa, H., Leung, J., Grimsey, N., Peak-Chew, S., Siniossoglou, S. (2005) The yeast lipin Smp2 couples phospholipid biosynthesis to nuclear membrane growth. EMBO J. 24, 1931–1941.
Sasser, T., Qiu, Q. S., Karunakaran, S., Padolina, M., Reyes, A., Flood, B., Smith, S., Gonzales, C., Fratti, R. A. (2012) Yeast lipin 1 orthologue pah1p regulates vacuole homeostasis and membrane fusion. J. Biol. Chem. 287, 2221–2236.
Schneiter, R., Daum, G. (2006) Extraction of yeast lipids. Methods Mol. Biol. 313, 41–45.
Sciorra, V. A., Morris, A. J. (2002) Roles for lipid phosphate phosphatases in regulation of cellular signaling. Biochim. Biophys. Acta 1582, 45–51.
Shahnazari, S., Yen, W. L., Birmingham, C. L., Shiu, J., Namolovan, A., Zheng, Y. T., Nakayama, K., Klionsky, D. J., Brumell, J. H. (2010) A diacylglycerol-dependent signaling pathway contributes to regulation of antibacterial autophagy. Cell Host Microbe 8, 137–146.
Siniossoglou, S. (2013) Phospholipid metabolism and nuclear function: roles of the lipin family of phosphatidic acid phosphatases. Biochim. Biophys. Acta 1831, 575–581.
Skinner, J. R., Shew, T. M., Schwartz, D. M., Tzekov, A., Lepus, C. M., Abumrad, N. A., Wolins, N. E. (2009) Diacylglycerol enrichment of endoplasmic reticulum or lipid droplets recruits perilipin 3/TIP47 during lipid storage and mobilization. J. Biol. Chem. 284, 30941–30948.
Suviolahti, E., Reue, K., Cantor, R. M., Phan, J., Gentile, M., Naukkarinen, J., Soro-Paavonen, A., Oksanen, L., Kaprio, J., Rissanen, A., Salomaa, V., Kontula, K., Taskinen, M. R., Pajukanta, P., Peltonen, L. (2006) Cross-species analyses implicate Lipin 1 involvement in human glucose metabolism. Hum. Mol. Genet. 15, 377–386.
Sztalryd, C., Xu, G., Dorward, H., Tansey, J. T., Contreras, J. A., Kimmel, A. R., Londos, C. (2003) Perilipin A is essential for the translocation of hormone-sensitive lipase during lipolytic activation. J. Cell Biol. 161, 1093–1103.
Wolinski, H., Kohlwein, S. D. (2008) Microscopic analysis of lipid droplet metabolism and dynamics in yeast. Methods Mol. Biol. 457, 151–163.
Zeharia, A., Shaag, A., Houtkooper, R. H., Hindi, T., De Lonlay, P., Erez, G., Hubert, L., Saada, A., De Keyzer, Y., Eshel, G., Vaz, F. M., Pines, O., Elpeleg, O. (2008) Mutations in LPIN1 cause recurrent acute myoglobinuria in childhood. Am. J. Hum. Genet. 83, 489–494.
Zimmermann, R., Strauss, J. G., Haemmerle, G., Schoiswohl, G., Birner-Gruenberger, R., Riederer, M., Lass, A., Neuberger, G., Eisenhaber, F., Hermetter, A., Zechner, R. (2004) Fat mobilization in adipose tissue is promoted by adipose triglyceride lipase. Science 306, 1383–1386.
Acknowledgements
We thank Dr. Ophry Pines and Orly Elpeleg for their kind gift of plasmid YEp-LPIN1. We are grateful to Dr. Deeksha Vishwamitra for her kind assistance in language proofing. This work was sponsored by grants from National Natural Science Foundation of China (31100549), the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry and Fundamental Research Funds for the Central Universities (2232014A3-03 and 222201313010), the National Special Fund for State Key Laboratory of Bioreactor Engineering (2060204).
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Fang, Z., Wang, S., Du, X. et al. Phosphatidate Phosphatase-1 is Functionally Conserved in Lipid Synthesis and Storage from Human to Yeast. BIOLOGIA FUTURA 65, 481–492 (2014). https://doi.org/10.1556/ABiol.65.2014.4.11
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DOI: https://doi.org/10.1556/ABiol.65.2014.4.11