A Genetic Screen for Mutants with Supersized Lipid Droplets in Caenorhabditis Elegans

Overview

Journal G3 (Bethesda)

Publisher Oxford University Press

Specialties Genetics
Molecular Biology

Date 2016 Jun 5

PMID 27261001

Citations 16

Authors

Shiwei Li

Shibin Xu

Yanli Ma

Shuang Wu

Yu Feng

Qingpo Cui

Lifeng Chen

Shuang Zhou

Yuanyuan Kong

Xiaoyu Zhang

Jialei Yu

Mengdi Wu

Shaobing O Zhang

Affiliations

Soon will be listed here.

Abstract

To identify genes that regulate the dynamics of lipid droplet (LD) size, we have used the genetically tractable model organism Caenorhabditis elegans, whose wild-type LD population displays a steady state of size with an upper limit of 3 μm in diameter. From a saturated forward genetic screen of 6.7 × 10(5) mutagenized haploid genomes, we isolated 118 mutants with supersized intestinal LDs often reaching 10 μm. These mutants define nine novel complementation groups, in addition to four known genes (maoc-1, dhs-28, daf-22, and prx-10). The nine groups are named drop (lipid droplet abnormal) and categorized into four classes. Class I mutants drop-5 and drop-9, similar to prx-10, are up-regulated in ACS-22-DGAT-2-dependent LD growth, resistant to LD hydrolysis, and defective in peroxisome import. Class II mutants drop-2, drop-3, drop-6, and drop-7 are up-regulated in LD growth, are resistant to LD hydrolysis, but are not defective in peroxisome import. Class III mutants drop-1 and drop-8 are neither up-regulated in LD growth nor resistant to LD hydrolysis, but seemingly up-regulated in LD fusion. Class IV mutant drop-4 is cloned as sams-1 and, different to the other three classes, is ACS-22-independent and hydrolysis-resistant. These four classes of supersized LD mutants should be valuable for mechanistic studies of LD cellular processes including growth, hydrolysis, and fusion.

Citing Articles

High Nutritional Conditions Influence Feeding Plasticity in Pristionchus pacificus and Render Worms Non-Predatory.

Piskobulu V, Athanasouli M, Witte H, Feldhaus C, Streit A, Sommer R J Exp Zool B Mol Dev Evol. 2025; 344(2):94-111.

PMID: 39822045 PMC: 11788882. DOI: 10.1002/jez.b.23284.

Differential roles of lysosomal cholesterol transporters in the development of NMJs.

Guo A, Wu Q, Yan X, Chen K, Liu Y, Liang D Life Sci Alliance. 2024; 7(10).

PMID: 39084875 PMC: 11291935. DOI: 10.26508/lsa.202402584.

Microbiota succession influences nematode physiology in a beetle microcosm ecosystem.

Lo W, Sommer R, Han Z Nat Commun. 2024; 15(1):5137.

PMID: 38879542 PMC: 11180206. DOI: 10.1038/s41467-024-49513-5.

4,4-Dimethylsterols Reduces Fat Accumulation via Inhibiting Fatty Acid Amide Hydrolase In Vitro and In Vivo.

Zhang T, Xie L, Guo Y, Wang Z, Guo X, Liu R Research (Wash D C). 2024; 7:0377.

PMID: 38812531 PMC: 11134202. DOI: 10.34133/research.0377.

Identification of genetic suppressors for a BSCL2 lipodystrophy pathogenic variant in Caenorhabditis elegans.

Bai X, Smith H, Golden A Dis Model Mech. 2024; 17(6).

PMID: 38454882 PMC: 11051982. DOI: 10.1242/dmm.050524.

References

Fei W, Shui G, Gaeta B, Du X, Kuerschner L, Li P . Fld1p, a functional homologue of human seipin, regulates the size of lipid droplets in yeast. J Cell Biol. 2008; 180(3):473-82. PMC: 2234226. DOI: 10.1083/jcb.200711136. View

Xu N, Zhang S, Cole R, McKinney S, Guo F, Haas J . The FATP1-DGAT2 complex facilitates lipid droplet expansion at the ER-lipid droplet interface. J Cell Biol. 2012; 198(5):895-911. PMC: 3432760. DOI: 10.1083/jcb.201201139. View

Zhang P, Na H, Liu Z, Zhang S, Xue P, Chen Y . Proteomic study and marker protein identification of Caenorhabditis elegans lipid droplets. Mol Cell Proteomics. 2012; 11(8):317-28. PMC: 3412964. DOI: 10.1074/mcp.M111.016345. View

Walker A, Jacobs R, Watts J, Rottiers V, Jiang K, Finnegan D . A conserved SREBP-1/phosphatidylcholine feedback circuit regulates lipogenesis in metazoans. Cell. 2011; 147(4):840-52. PMC: 3384509. DOI: 10.1016/j.cell.2011.09.045. View

Soukas A, Carr C, Ruvkun G . Genetic regulation of Caenorhabditis elegans lysosome related organelle function. PLoS Genet. 2013; 9(10):e1003908. PMC: 3812091. DOI: 10.1371/journal.pgen.1003908. View

Szymanski K, Binns D, Bartz R, Grishin N, Li W, Agarwal A . The lipodystrophy protein seipin is found at endoplasmic reticulum lipid droplet junctions and is important for droplet morphology. Proc Natl Acad Sci U S A. 2007; 104(52):20890-5. PMC: 2409237. DOI: 10.1073/pnas.0704154104. View

Liang B, Ferguson K, Kadyk L, Watts J . The role of nuclear receptor NHR-64 in fat storage regulation in Caenorhabditis elegans. PLoS One. 2010; 5(3):e9869. PMC: 2845610. DOI: 10.1371/journal.pone.0009869. View

Zhang S, Trimble R, Guo F, Mak H . Lipid droplets as ubiquitous fat storage organelles in C. elegans. BMC Cell Biol. 2010; 11:96. PMC: 3004847. DOI: 10.1186/1471-2121-11-96. View

Klemm R, Norton J, Cole R, Li C, Park S, Crane M . A conserved role for atlastin GTPases in regulating lipid droplet size. Cell Rep. 2013; 3(5):1465-75. PMC: 3742324. DOI: 10.1016/j.celrep.2013.04.015. View

10.

Zhang S, Box A, Xu N, Le Men J, Yu J, Guo F . Genetic and dietary regulation of lipid droplet expansion in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 2010; 107(10):4640-5. PMC: 2842062. DOI: 10.1073/pnas.0912308107. View

11.

Brenner S . The genetics of Caenorhabditis elegans. Genetics. 1974; 77(1):71-94. PMC: 1213120. DOI: 10.1093/genetics/77.1.71. View

12.

Anderson P . Mutagenesis. Methods Cell Biol. 1995; 48:31-58. View

13.

De Stasio E, Dorman S . Optimization of ENU mutagenesis of Caenorhabditis elegans. Mutat Res. 2001; 495(1-2):81-8. DOI: 10.1016/s1383-5718(01)00198-x. View

14.

ORourke E, Soukas A, Carr C, Ruvkun G . C. elegans major fats are stored in vesicles distinct from lysosome-related organelles. Cell Metab. 2009; 10(5):430-5. PMC: 2921818. DOI: 10.1016/j.cmet.2009.10.002. View

15.

Ploegh H . A lipid-based model for the creation of an escape hatch from the endoplasmic reticulum. Nature. 2007; 448(7152):435-8. DOI: 10.1038/nature06004. View

16.

Brooks K, Liang B, Watts J . The influence of bacterial diet on fat storage in C. elegans. PLoS One. 2009; 4(10):e7545. PMC: 2760100. DOI: 10.1371/journal.pone.0007545. View

17.

Butcher R, Ragains J, Li W, Ruvkun G, Clardy J, Mak H . Biosynthesis of the Caenorhabditis elegans dauer pheromone. Proc Natl Acad Sci U S A. 2009; 106(6):1875-9. PMC: 2631283. DOI: 10.1073/pnas.0810338106. View

18.

Sato K, Ernstrom G, Watanabe S, Weimer R, Chen C, Sato M . Differential requirements for clathrin in receptor-mediated endocytosis and maintenance of synaptic vesicle pools. Proc Natl Acad Sci U S A. 2009; 106(4):1139-44. PMC: 2633560. DOI: 10.1073/pnas.0809541106. View

19.

Liu Z, Li X, Ge Q, Ding M, Huang X . A lipid droplet-associated GFP reporter-based screen identifies new fat storage regulators in C. elegans. J Genet Genomics. 2014; 41(5):305-13. DOI: 10.1016/j.jgg.2014.03.002. View

20.

Klapper M, Ehmke M, Palgunow D, Bohme M, Matthaus C, Bergner G . Fluorescence-based fixative and vital staining of lipid droplets in Caenorhabditis elegans reveal fat stores using microscopy and flow cytometry approaches. J Lipid Res. 2011; 52(6):1281-1293. PMC: 3090249. DOI: 10.1194/jlr.D011940. View