Activation of de novo lipogenesis in cancer cells is increasingly recognized as a hallmark of aggressive cancers and has been implicated in the production of membranes for rapid cell proliferation. In the current report, we provide evidence that this activation has a more profound role. Using a mass spectrometry-based phospholipid analysis approach, we show that clinical tumor tissues that display the lipogenic phenotype show an increase in the degree of lipid saturation compared with nonlipogenic tumors. Reversal of the lipogenic switch in cancer cells by treatment with the lipogenesis inhibitor soraphen A or by targeting lipogenic enzymes with small interfering RNA leads to a marked decrease in saturated and mono-unsaturated phospholipid species and increases the relative degree of polyunsaturation. Because polyunsaturated acyl chains are more susceptible to peroxidation, inhibition of lipogenesis increases the levels of peroxidation end products and renders cells more susceptible to oxidative stress-induced cell death. As saturated lipids pack more densely, modulation of lipogenesis also alters lateral and transversal membrane dynamics as revealed by diffusion of membrane-targeted green fluorescent protein and by the uptake and response to doxorubicin. These data show that shifting lipid acquisition from lipid uptake toward de novo lipogenesis dramatically changes membrane properties and protects cells from both endogenous and exogenous insults. These findings provide important new insights into the role of de novo lipogenesis in cancer cells, and they provide a rationale for the use of lipogenesis inhibitors as antineoplastic agents and as chemotherapeutic sensitizers. Cancer Res; 70(20); 8117-26. ©2010 AACR.

γ-Secretases are a family of intramembrane-cleaving proteases involved in various signaling pathways and diseases, including Alzheimer's disease (AD). Cells co-express differing γ-secretase complexes, including two homologous presenilins (PSENs). We examined the significance of this heterogeneity and identified a unique motif in PSEN2 that directs this γ-secretase to late endosomes/lysosomes via a phosphorylation-dependent interaction with the AP-1 adaptor complex. Accordingly, PSEN2 selectively cleaves late endosomal/lysosomal localized substrates and generates the prominent pool of intracellular Aβ that contains longer Aβ; familial AD (FAD)-associated mutations in PSEN2 increased the levels of longer Aβ further. Moreover, a subset of FAD mutants in PSEN1, normally more broadly distributed in the cell, phenocopies PSEN2 and shifts its localization to late endosomes/lysosomes. Thus, localization of γ-secretases determines substrate specificity, while FAD-causing mutations strongly enhance accumulation of aggregation-prone Aβ42 in intracellular acidic compartments. The findings reveal potentially important roles for specific intracellular, localized reactions contributing to AD pathogenesis.

Zinc co-crystallizes with insulin in dense core secretory granules, but its role in insulin biosynthesis, storage and secretion is unknown. In this study we assessed the role of the zinc transporter ZnT8 using ZnT8-knockout (ZnT8 ؊/؊ ) mice. Absence of ZnT8 expression caused loss of zinc release upon stimulation of exocytosis, but normal rates of insulin biosynthesis, normal insulin content and preserved glucose-induced insulin release. Ultrastructurally, mature dense core insulin granules were rare in ZnT8 ؊/؊ beta cells and were replaced by immature, pale insulin ''progranules,'' which were larger than in ZnT8 ؉/؉ islets. When mice were fed a control diet, glucose tolerance and insulin sensitivity were normal. However, after high-fat diet feeding, the ZnT8 ؊/؊ mice became glucose intolerant or diabetic, and islets became less responsive to glucose. Our data show that the ZnT8 transporter is essential for the formation of insulin crystals in beta cells, contributing to the packaging efficiency of stored insulin. Interaction between the ZnT8 ؊/؊ genotype and diet to induce diabetes is a model for further studies of the mechanism of disease of human ZNT8 gene mutations.dense core granule ͉ diabetes ͉ zinc

UFM1 is a member of the ubiquitin like protein family. While the enzymatic cascade of UFM1 conjugation has been elucidated in recent years, the biological function remains largely unknown. In this report we demonstrate that the recently identified C20orf116 [1], which we name UFM1-binding protein 1 containing a PCI domain (UFBP1), andCDK5RAP3 interact with UFM1. Components of the UFM1 conjugation pathway (UFM1, UFBP1, UFL1 and CDK5RAP3) are highly expressed in pancreatic islets of Langerhans and some other secretory tissues. Co-localization of UFM1 with UFBP1 in the endoplasmic reticulum (ER)depends on UFBP1. We demonstrate that ER stress, which is common in secretory cells, induces expression of Ufm1, Ufbp1 and Ufl1 in the beta-cell line INS-1E.siRNA-mediated Ufm1 or Ufbp1knockdown enhances apoptosis upon ER stress.Silencing the E3 enzyme UFL1, results in similar outcomes, suggesting that UFM1-UFBP1 conjugation is required to prevent ER stress-induced apoptosis. Together, our data suggest that UFM1-UFBP1participate in preventing ER stress-induced apoptosis in protein secretory cells.

In response to crowding, locusts develop characteristic black patterns that are well discernible in the gregarious phase at outbreaks. We report here a dark-colorinducing neuropeptide (dark-pigmentotropin) from the corpora cardiaca of two plague locusts, Schistocerca gregaria and Locusta migratoria. The chromatographic isolation of this neuropeptide was monitored by using a bioassay with an albino mutant of L. migratoria. Body-color polymorphism is widespread among animals. Two locust species, Schistocerca gregaria and Locusta migratoria, display conspicuous differences in body color, particularly during the nymphal stage. A major extrinsic factor influencing locust body color is phase polymorphism, a term used to describe continuous polymorphism in response to population density: locusts at a low density (solitary phase) are often green or brown, whereas those at outbreaks (gregarious phase) develop black patterns (1-4). Although the role of juvenile hormone in the induction of the green color is well established (2-4), little information is available about the hormonal factor that induces dark color in locusts. It has long been known that some factor present in the brain and the corp cardiaca (CC) promotes darkening in locusts (2, 3), but progress in identifying its chemical nature has been hampered by the lack of a convenient bioassay.Recently, we discovered an albino mutant, originating from a laboratory colony of an Okinawa (Japan) strain of L. migratoria (5). Albinism in this mutant is controlled by a single recessive Mendelian unite (5), as described also for other albino mutants of this species (6, 7), as well as of S. gregaria (8) and the grasshopper Melanoplus sanguinipes (under the name Melanoplus bilituratus) (9). The albinism in the Okinawa strain of L. migratoria is caused by the deficiency of a peptide(s) present in the central nervous system and the CC. Implantation of a brain or CC taken from normal (pigmented) individuals or injection of their methanolic extract induces dark color in albino locusts (10-12), but injection of such methanolic extract made from albino individuals has no dark-colorinducing effect in albino locusts (11). Of interest, implantation of brains or CC taken from other taxa, including S. gregaria and other acridids, cockroaches, katydids, crickets, and moths also are effective in inducing dark color in albino L. migratoria (10,12,13). This result indicates that similar substances inducing dark color in L. migratoria may exist in diverse groups of insects. Because whitish albino locusts can be obtained easily by mass rearing, they provide an excellent bioassay system for the characterization of this dark-color inducing peptide. Its role in body color polymorphism and phase polymorphism in locusts can then be determined by means of the synthetic analog. MATERIALS AND METHODSInsects and Tissue Extraction. The colony of the desert locust S. gregaria was maintained according to Ashby's method (14) and that of the migratory locust L. migratoria migratorioides as described...