Lipid composition of <i>Spodoptera frugiperda</i> (Sf9) and <i>Trichoplusia ni</i> (Tn) insect cells used for baculovirus infection

Marheineke, Kathrin; Grünewald, Sylvia; Christie, William W.; Reiländer, Helmut

doi:10.1016/s0014-5793(98)01523-3

Cited by 148 publications

(120 citation statements)

References 20 publications

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“…Our data largely agree with published studies analyzing the lipid composition of insect cell membranes (11), in particular SF9 cells (12), especially regarding limited amounts of cholesterol as well as for their high PE content. In contrast, we found that SF9 similarly to HEK 293T cells contain all major PL species, notably PG, PS, and SM, which was described for Drosoph- (13).…”

Section: Lipidic Composition Of Insect and Mammaliansupporting

confidence: 92%

Phosphatidylethanolamine Is a Key Regulator of Membrane Fluidity in Eukaryotic Cells

Dawaliby

Trubbia

Delporte

et al. 2016

Journal of Biological Chemistry

306

202

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Adequate membrane fluidity is required for a variety of key cellular processes and in particular for proper function of membrane proteins. In most eukaryotic cells, membrane fluidity is known to be regulated by fatty acid desaturation and cholesterol, although some cells, such as insect cells, are almost devoid of sterol synthesis. We show here that insect and mammalian cells present similar microviscosity at their respective physiological temperature. To investigate how both sterols and phospholipids control fluidity homeostasis, we quantified the lipidic composition of insect SF9 and mammalian HEK 293T cells under normal or sterol-modified condition. As expected, insect cells show minimal sterols compared with mammalian cells. A major difference is also observed in phospholipid content as the ratio of phosphatidylethanolamine (PE) to phosphatidylcholine (PC) is inverted (4 times higher in SF9 cells). In vitro studies in liposomes confirm that both cholesterol and PE can increase rigidity of the bilayer, suggesting that both can be used by cells to maintain membrane fluidity. We then show that exogenously increasing the cholesterol amount in SF9 membranes leads to a significant decrease in PE:PC ratio whereas decreasing cholesterol in HEK 293T cells using statin treatment leads to an increase in the PE:PC ratio. In all cases, the membrane fluidity is maintained, indicating that both cell types combine regulation by sterols and phospholipids to control proper membrane fluidity.

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Section: Lipidic Composition Of Insect and Mammaliansupporting

confidence: 92%

Phosphatidylethanolamine Is a Key Regulator of Membrane Fluidity in Eukaryotic Cells

Dawaliby

Trubbia

Delporte

et al. 2016

Journal of Biological Chemistry

306

202

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“…Thus, viral envelopes are likely to contain more PE than PS. With PE content in insect cells being even higher (as much as 46-58%) (44)(45)(46)(47)(48), it is not surprising that arboviruses, such as flavi-and alphaviruses, evolved to use PE-binding receptors like TIM1 with high efficiency (9,10).…”

Section: Discussionmentioning

confidence: 99%

Virion-associated phosphatidylethanolamine promotes TIM1-mediated infection by Ebola, dengue, and West Nile viruses

Richard

Zhang

Park

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

119

105

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Phosphatidylserine (PS) receptors contribute to two crucial biological processes: apoptotic clearance and entry of many enveloped viruses. In both cases, they recognize PS exposed on the plasma membrane. Here we demonstrate that phosphatidylethanolamine (PE) is also a ligand for PS receptors and that this phospholipid mediates phagocytosis and viral entry. We show that a subset of PS receptors, including T-cell immunoglobulin (Ig) mucin domain protein 1 (TIM1), efficiently bind PE. We further show that PE is present in the virions of flaviviruses and filoviruses, and that the PE-specific cyclic peptide lantibiotic agent Duramycin efficiently inhibits the entry of West Nile, dengue, and Ebola viruses. The inhibitory effect of Duramycin is specific: it inhibits TIM1-mediated, but not L-SIGNmediated, virus infection, and it does so by blocking virus attachment to TIM1. We further demonstrate that PE is exposed on the surface of apoptotic cells, and promotes their phagocytic uptake by TIM1-expressing cells. Together, our data show that PE plays a key role in TIM1-mediated virus entry, suggest that disrupting PE association with PS receptors is a promising broad-spectrum antiviral strategy, and deepen our understanding of the process by which apoptotic cells are cleared. M embers of the filovirus and flavivirus families are the causative agents of life-threatening diseases. Ebola virus (EBOV), a filovirus, causes hemorrhagic fever with an average case fatality rate as high as 65% (1). Although there are EBOV vaccine candidates (2, 3), there is currently no licensed vaccine or treatment. Dengue virus (DENV) and West Nile virus (WNV) belong to the flavivirus family. Both are transmitted to humans through mosquito bites and can cause lethal hemorrhagic fever (in the case of DENV) or severe neurological diseases (in the case of WNV) (4, 5). Flaviviruses are emerging as major health concerns in tropical and subtropical areas worldwide. More than one third of the world's population is estimated to be at risk for DENV infection, with approximately 400 million people infected yearly (6). There are currently no approved vaccines or therapeutic agents against DENV or WNV.Virus entry into host cells typically initiates with the interaction between viral entry glycoproteins (GPs) and a receptor or coreceptor expressed at the surface of the target cell. Viruses also use less specific mechanisms to localize to target cell membranes, for example through GP association with various attachment factors (7). During the past few years, it has been increasingly recognized that many viruses also use a strategy known as apoptotic mimicry to promote their association with, and internalization into their target cells (8). Receptors for phospholipids, specifically phosphatidylserine (PS), normally involved in the clearance of apoptotic cells, markedly enhance the infection of a number of enveloped viruses. These PS receptors are presumed to engage PS on the virion membrane rather than the viral entry protein (9, 10). Enveloped viruses acquire...

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“…This observation has been explained in terms of an altered permeability of these cells combined with the ability of ribotoxins to interact with acid phospholipid-containing membranes (Gasset et al, 1989(Gasset et al, , 1990Herrero-Galán et al, 2008, 2012bMartínez-Ruiz et al, 2001;Olmo et al, 2001). Insect cells have a different plasma membrane composition from mammalian cells due to their higher content of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol and a significant lower cholesterol/phospholipid ratio (Marheineke et al, 1998). Therefore insect plasma membranes are thinner and probably more fluid than those in mammalian cells being better candidates as targets for ribotoxins.…”

Section: Discussionmentioning

confidence: 99%

Fungal extracellular ribotoxins as insecticidal agents

Olombrada¹,

Herrero-Galán²,

Tello³

et al. 2013

Insect Biochemistry and Molecular Biology

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Fungal ribotoxins were discovered almost 50 years ago as extracellular ribonucleases (RNases) with antitumoral properties. However, the biological function of these toxic proteins has remained elusive. The discovery of the ribotoxin HtA, produced by the invertebrates pathogen H. thompsonii, revived the old proposal that insecticidal activity would be their long searched function. Unfortunately, HtA is rather singular among all ribotoxins known in terms of sequence and structure similarities. Thus, it was intriguing to answer the question of whether HtA is just an exception or, on the contrary, the paradigmatic example of the ribotoxins function. The work presented uses HtA and -sarcin, the most representative member of the ribotoxins family, to show their strong toxic action against insect larvae and cells.

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Lipid composition of Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn) insect cells used for baculovirus infection

Cited by 148 publications

References 20 publications

Phosphatidylethanolamine Is a Key Regulator of Membrane Fluidity in Eukaryotic Cells

Phosphatidylethanolamine Is a Key Regulator of Membrane Fluidity in Eukaryotic Cells

Virion-associated phosphatidylethanolamine promotes TIM1-mediated infection by Ebola, dengue, and West Nile viruses

Fungal extracellular ribotoxins as insecticidal agents

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