Pathways of glutamine metabolism in Spodoptera frugiperda (Sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1H/15N NMR

Drews, Monika; Doverskog, Magnus; Öhman, Lars; Chapman, Bogdan E.; Jacobsson, Ulla; Kuchel, Philip W.; Häggström, Lena

doi:10.1016/s0168-1656(99)00231-x

Cited by 56 publications

(61 citation statements)

References 33 publications

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“…GDH in body tissues is likely involved in generation of ammonia from glutamate, the activity considered as being usual in animal tissues. GS/GltS is the dominant mode of ammonia assimilation when glucose is plentiful (because GS is ATP-dependent) and ammonia is in low concentration (27)(28)(29). These conditions apply to aphids, which ingest excess sugar and must maintain ammonia at low concentrations in cells because of its toxicity.…”

Section: And Table 1)mentioning

confidence: 99%

Aphid genome expression reveals host–symbiont cooperation in the production of amino acids

Hansen

Moran

2011

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

365

478

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The evolution of intimate symbiosis requires the coordination of gene expression and content between the distinct partner genomes; this coordination allows the fusion of capabilities of each organism into a single integrated metabolism. In aphids, the 10 essential amino acids are scarce in the phloem sap diet and are supplied by the obligate bacterial endosymbiont ( Buchnera ), which lives inside specialized cells called bacteriocytes. Although Buchnera ’s genome encodes most genes for essential amino acid biosynthesis, several genes in essential amino acid pathways are missing, as are most genes for production of nonessential amino acids. Additionally, it is unresolved whether the supply of nitrogen for amino acid biosynthesis is supplemented by recycling of waste ammonia. We compared pea aphid gene expression between bacteriocytes and other body tissues using RNA sequencing and pathway analysis and exploiting the genome sequences available for both partners. We found that 26 genes underlying amino acid biosynthesis were up-regulated in bacteriocytes. Seven of these up-regulated genes fill the gaps of Buchnera ’s essential amino acid pathways. In addition, genes underlying five nonessential amino acid pathways lost from Buchnera are up-regulated in bacteriocytes. Finally, our results reveal that two genes, glutamine synthetase and glutamate synthase, which potentially work together in the incorporation of ammonium nitrogen into glutamate (GOGAT) cycle to assimilate ammonia into glutamate, are up-regulated in bacteriocytes. Thus, host gene expression and symbiont capabilities are closely integrated within bacteriocytes, which function as specialized organs of amino acid production. Furthermore, the GOGAT cycle may be a key source of nitrogen fueling the integrated amino acid metabolism of the aphid– Buchnera partnership.

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Section: And Table 1)mentioning

confidence: 99%

Aphid genome expression reveals host–symbiont cooperation in the production of amino acids

Hansen

Moran

2011

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

365

478

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“…In this respect, DSIR-HA-1179 cells share a similarity with the metabolic nature of the Sf-9 cell line. It would be well worth investigating the production of ethanol and glycerol which have been identified as by-products of glucose metabolism in the Sf-9 cell line (Drews et al 2000). In addition, in order to gain a complete understanding into the metabolism of the cell line, detailed amino acid analyses must be performed.…”

Section: Metabolites Analysesmentioning

confidence: 99%

“…Glucose has been found to be the major carbohydrate source in a range of insect cell lines including Sf-9 (Reuveny et al 1992), HighFive (Rhiel et al 1997), saUFL-Ag-286 (Gioria et al 2006) and BCIRL-HZ-AM1 (Lua and Reid 2003). BTI-Tn-5B1-4 and BCIRl-HZ-AM1 cells display an inherent tendency to produce lactate and ammonia even when provided with sufficient glucose to grow in batch culture (Rhiel et al 1997;Lua and Reid 2003), while Sf-9 cells will produce them only under glucose deprivation (Ohman et al 1995;Drews et al 2000). On the other hand, the metabolism of the saUFL-AG-286 cell line is characterized by the production of ammonia but not of lactate (Gioria et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Characterization of growth and Oryctes rhinoceros nudivirus production in attached cultures of the DSIR-HA-1179 coleopteran insect cell line

et al. 2013

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The DSIR-HA-1179 coleopteran cell line is a susceptible and permissive host to the Oryctes rhinoceros nudivirus (OrNV), which has been used as a biocontrol agent against the coconut rhinoceros beetle (Oryctes rhinoceros); a pest of palms in the Asia-Pacific region. However, little is known about growth and metabolism of this cell line, knowledge of which is necessary to develop an in vitro large-scale OrNV production process. The strong anchoragedependent characteristics of the cell line, its particular fragility and its tendency to form dense clumps when manipulated, are the most likely reasons that have precluded further development of the cell line. In order to characterize DSIR-HA-1179 cells, there was first a need for a reliable technique to count the cells. A homogenous cell suspension suitable for enumeration could be produced by treatment with TrypLE Express TM with optimum mean time for cell release calculated as 30 min. The cell line was adapted to grow in four serum-supplemented culture media namely TC-100, IPL-41, Sf-900 II and Sf-900 III and cell growth, glucose consumption, lactate and ammonia production were assessed from static-batch cultures. The maximum viable cell density was reached in Sf-900 II (17.9 9 10 5 cells/ml), with the maximum specific growth rate observed in this culture medium as well (0.0074 h -1 ). Higher production of OrNV was observed in IPL-41 and TC-100 (4.1 9 10 7 TCID 50 /ml) than in cultures infected in Sf-900 III (2.0 9 10 7 TCID 50 /ml) and Sf-900 II (1.4 9 10 7 TCID 50 /ml). At the end of the growth period, glucose was completely consumed in cultures grown in TC-100, while remained in excess in the other three culture media. The cell line produced lactate and ammonia to very low levels in the TC-100 culture medium which is a promising aspect for its cultivation at large-scale.

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“…However, these results are still controversial, because other studies have found that the percentage of glucose consumed that is derived to the tricarboxylic acid cycle is much lower (Benslimane et al, 2005). The production of other metabolites such as alanine, glycerol and ethanol, as well as fatty acid synthesis, could explain the fate of a significant fraction of carbon incorporated to Sf9 cells through glucose consumption (Drews et al, 2000;Bernal et al, 2009). BTI-TN-5B1-4 and BM5 cell lines have a different behavior: both cell lines produce lactate, displaying a behavior similar to that of mammalian cells (Rhiel et al, 1997;Stavroulakis et al, 1991).…”

Section: Nutrition and Metabolism Of Lepidopteran Insect Cellsmentioning

confidence: 99%

“…Besides serving as precursors for the synthesis of proteins and nucleic acids, amino acids that are consumed faster are used as sources of energy, such as glutamine and asparagine. The metabolism of glutamine has been studied in Sf9 cells, where the utilization pathways are depending on the availability of glucose (Drews et al, 2000). It has been proposed that, in glucose excess, the cytoplasmatic enzyme glutamate synthase transfers the amidic nitrogen of glutamine to the amine-position in glutamate, from where it is transaminated to alanine, the main product of glutamine metabolism.…”

Section: Nutrition and Metabolism Of Lepidopteran Insect Cellsmentioning

confidence: 99%

Production of Insecticidal Baculoviruses in Insect Cell Cultures: Potential and Limitations

Claus¹,

Gioria²,

Micheloud³

et al. 2012

Insecticides - Basic and Other Applications

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Pathways of glutamine metabolism in Spodoptera frugiperda (Sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1H/15N NMR

Cited by 56 publications

References 33 publications

Aphid genome expression reveals host–symbiont cooperation in the production of amino acids

Aphid genome expression reveals host–symbiont cooperation in the production of amino acids

Characterization of growth and Oryctes rhinoceros nudivirus production in attached cultures of the DSIR-HA-1179 coleopteran insect cell line

Production of Insecticidal Baculoviruses in Insect Cell Cultures: Potential and Limitations

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