Inactivation of the C-elegans lipin homolog leads to ER disorganization and to defects in the breakdown and reassembly of the nuclear envelope

The Laboratory of Biochemistry and Genetics and National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 8 Center Drive, Bethesda, MD 20892, USA.
Journal of Cell Science (Impact Factor: 5.43). 07/2009; 122(Pt 12):1970-8. DOI: 10.1242/jcs.044743
Source: PubMed


The nuclear envelope (NE) is a dynamic structure, undergoing periods of growth, breakdown and reassembly during the cell cycle. In yeast, altering lipid synthesis by inactivating the yeast homolog of lipin, a phosphatidic acid phosphohydrolase, leads to disorganization of the peripheral ER and abnormal nuclear shape. These results suggest that lipid metabolism contributes to NE dynamics; however, since yeast undergo closed mitosis, the relevance of these observations to higher eukaryotes is unclear. In mammals, lipin has been implicated in adipose tissue differentiation, insulin resistance, lipid storage and obesity, but the underlying cellular defects caused by altering lipin levels are not known. Here, we identify the Caenorhabditis elegans lipin homolog (LPIN-1) and examine its affect on NE dynamics. We find that downregulating LPIN-1 by RNAi results in the appearance of membrane sheets and other abnormal structures in the peripheral ER. Moreover, lpin-1 RNAi causes defects in NE breakdown, abnormal chromosome segregation and irregular nuclear morphology. These results uncover cellular processes affected by lipin in metazoa, and suggest that lipid synthesis has a role in NE dynamics.

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    • "RIBO-1, for example, has been shown to play a role in chromosome segregation [52]. The polarized ER aggregations observed in OSTD-1 depletions are also distinct from the distribution of ER aggregates reported in ribo-1, car-1 and lpin-1 RNAi-treated embryos or the more disperse distribution of ER in rab-5 and yop-1/ret-1 depletions [59,61,81], suggesting that OSTD-1 may be involved in maintaining ER morphology specifically in the anterior. It is not clear how the polarization of ER functions during mitosis, but transitional ER (tER) sites have been shown to be polarized in the hyphal tips of fungi where growth and morphogenesis occur [82]. "
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    ABSTRACT: Cell division is important for many cellular processes including cell growth, reproduction, wound healing and stem cell renewal. Failures in cell division can often lead to tumors and birth defects. To identify factors necessary for this process, we implemented a comparative profiling strategy of the published mitotic spindle proteome from our laboratory. Of the candidate mammalian proteins, we determined that 77% had orthologs in Caenorhabditis elegans and 18% were associated with human disease. Of the C. elegans candidates (n=146), we determined that 34 genes functioned in embryonic development and 56% of these were predicted to be membrane trafficking proteins. A secondary, visual screen to detect distinct defects in cell division revealed 21 genes that were necessary for cytokinesis. One of these candidates, OSTD-1, an ER resident protein, was further characterized due to the aberrant cleavage furrow placement and failures in division. We determined that OSTD-1 plays a role in maintaining the dynamic morphology of the ER during the cell cycle. In addition, 65% of all ostd-1 RNAi-treated embryos failed to correctly position cleavage furrows, suggesting that proper ER morphology plays a necessary function during animal cell division.
    PLoS ONE 10/2013; 8(10):e77051. DOI:10.1371/journal.pone.0077051 · 3.23 Impact Factor
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    • "Clinicians should be aware that a defective heart function due to intracardiac accumulation of lipids should be considered in this disease [48]. Because DAG, the product of PAP1 activity, is also the precursor of several membrane phospholipids [49] [50], altered cell membrane composition and hence increased fragility or sensitivity to breakdown injuries have been hypothesized [2]. However, skeletal muscle and myoblast phospholipid contents appeared unchanged, and proinflammatory stimuli did not significantly affect phospholipid metabolism in the myoblasts. "
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    ABSTRACT: Lipin-1 deficiency is associated with massive rhabdomyolysis episodes in humans, precipitated by febrile illnesses. Despite well-known roles of lipin-1 in lipid biosynthesis and transcriptional regulation, the pathogenic mechanisms leading to rhabdomyolysis remain unknown. Here we show that primary myoblasts from lipin-1-deficient patients exhibit a dramatic decrease in LPIN1 expression and phosphatidic acid phosphatase 1 activity, and a significant accumulation of lipid droplets (LD). The expression levels of LPIN1-target genes (PPARδ, PPARα, PGC-1α, ACADVL, CPT1B and CPT2) were not affected while lipin-2 protein level, a closely related member of the family, was increased. Microarray analysis of patients' myotubes identified 19 down-regulated and 51 up-regulated genes, indicating pleiotropic effects of lipin-1 deficiency. Special attention was paid to the up-regulated ACACB (acetyl-CoA carboxylase beta), a key enzyme in the fatty acid synthesis/oxidation balance. We demonstrated that overexpression of ACACB was associated with free fatty acid accumulation in patients' myoblasts whereas malonylcarnitine (as a measure of malonyl-CoA) and CPT1 activity were in the normal range in basal conditions accordingly to the normal daily activity reported by the patients. Remarkably ACACB invalidation in patients' myoblasts decreased LD number and size while LPIN1 invalidation in controls induced LD accumulation. Further, pro-inflammatory treatments (TNF1α+IL-1ß) designed to mimic febrile illness, resulted in increased malonylcarnitine levels, reduced CPT1 activity and enhanced LD accumulation, a phenomenon reversed by dexamethasone and TNFα or IL1-ß inhibitors. Our data suggest that the pathogenic mechanism of rhabdomyolysis in lipin-1-deficient patients combines the predisposing constitutive impairment of lipid metabolism and its exacerbation by pro-inflammatory cytokines.
    Biochimica et Biophysica Acta 08/2013; 1832(12). DOI:10.1016/j.bbadis.2013.07.021 · 4.66 Impact Factor
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    • "In humans and mice, Lpin-1 is associated with metabolic syndrome and type 2 diabetes [77-80] and mutations in human LPIN2 cause Majeed syndrome [81]. C. elegans lpin-1 has been implicated in affecting fat storage and the breakdown and assembly of the nuclear envelope [54,55]. Consistent with the mammalian Lpin1 mutation, inactivation of C. elegans lpin-1 by RNAi displayed resulted in low fat with small size of lipid droplets, as indicted by Nile Red staining (Figure 4). "
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    ABSTRACT: Background Animal models are indispensable to understand the lipid metabolism and lipid metabolic diseases. Over the last decade, the nematode Caenorhabditis elegans has become a popular animal model for exploring the regulation of lipid metabolism, obesity, and obese-related diseases. However, the genomic and functional conservation of lipid metabolism from C. elegans to humans remains unknown. In the present study, we systematically analyzed genes involved in lipid metabolism in the C. elegans genome using comparative genomics. Results We built a database containing 471 lipid genes from the C. elegans genome, and then assigned most of lipid genes into 16 different lipid metabolic pathways that were integrated into a network. Over 70% of C. elegans lipid genes have human orthologs, with 237 of 471 C. elegans lipid genes being conserved in humans, mice, rats, and Drosophila, of which 71 genes are specifically related to human metabolic diseases. Moreover, RNA-mediated interference (RNAi) was used to disrupt the expression of 356 of 471 lipid genes with available RNAi clones. We found that 21 genes strongly affect fat storage, development, reproduction, and other visible phenotypes, 6 of which have not previously been implicated in the regulation of fat metabolism and other phenotypes. Conclusions This study provides the first systematic genomic insight into lipid metabolism in C. elegans, supporting the use of C. elegans as an increasingly prominent model in the study of metabolic diseases.
    BMC Genomics 03/2013; 14(1):164. DOI:10.1186/1471-2164-14-164 · 3.99 Impact Factor
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