02558nas a2200193   4500008004100000245014400041210006900185260000800254300001200262490000700274520180700281100003802088700002602126700001602152700001902168700003102187700001902218856012702237       2017                        eng d00aCHO Cells Adapted to Inorganic Phosphate Limitation Show Higher Growth and Higher Pyruvate Carboxylase Flux in Phosphate Replete Conditions0 aCHO Cells Adapted to Inorganic Phosphate Limitation Show Higher   cMAY  a749-7580 v333 aInorganic phosphate (P-i) is an essential ion involved in diverse cellular processes including metabolism. Changes in cellular metabolism upon long term adaptation to P-i limitation have been reported in E. coli. Given the essential role of P-i, adaptation to P-i limitation may also result in metabolic changes in animal cells. In this study, we have adapted CHO cells producing recombinant IgG to limiting P-i conditions for 75 days. Not surprisingly, adapted cells showed better survival under P-i limitation. Here, we report the finding that such cells also showed better growth characteristics compared to control in batch culture replete with P-i ( higher peak density and integral viable cell density), accompanied by a lower specific oxygen uptake rate and cytochrome oxidase activity towards the end of exponential phase. Surprisingly, the adapted cells grew to a lower peak density under glucose limitation. This suggests long term P-i limitation may lead to selection for an altered metabolism with higher dependence on glucose availability for biomass assimilation compared to control. Steady state U-C-13 glucose labeling experiments suggest that adapted cells have a higher pyruvate carboxylase flux. Consistent with this observation, supplementation with aspartate abolished the peak density difference whereas supplementation with serine did not abolish the difference. This supports the hypothesis that cell growth in the adapted culture might be higher due to a higher pyruvate carboxylase flux. Decreased fitness under carbon limitation and mutations in the sucABCD operon has been previously reported in E. coli upon long term adaptation to P-i limitation, suggestive of a similarity in cellular response among such diverse species. (C) 2017 American Institute of Chemical Engineers1 aMaralingannavar, Vishwanathgouda;1 aParmar, Dharmeshkumar1 aPant, Tejal1 aGadgil, Chetan1 aPanchagnula, Venkateswarlu1 aGadgil, Mugdha  uhttps://library.ncl.res.in/content/cho-cells-adapted-inorganic-phosphate-limitation-show-higher-growth-and-higher-pyruvate01845nas a2200193   4500008004100000022001400041245013700055210006900192260000900261300001200270490000700282520115600289100000501445700001901450700002001469700001801489700002301507856012101530       2019                        eng d  a2213-671100aClathrin-mediated endocytosis regulates a balance between opposing signals to maintain the pluripotent state of embryonic stem cells0 aClathrinmediated endocytosis regulates a balance between opposin  cJAN   a152-1640 v123 a<p>Endocytosis is implicated in the maintenance of embryonic stem cell (ESC) pluripotency, although its exact role and the identity of molecular players remain poorly understood. Here, we show that the clathrin heavy chain (CLTC), involved in clathrin-mediated endocytosis (CME), is vital for maintaining mouse ESC (mESC) pluripotency. Knockdown of Cltc resulted in a loss of pluripotency accompanied by reduced E-cadherin (E-CAD) levels and increased levels of transforming growth factor beta (TGF-beta) and extracellular signal-regulated kinase (ERK) signaling. We demonstrate that both E-CAD and TGF-beta receptor type 1 (TGF-beta R1) are internalized through CME in mESCs. While E-CAD is recycled, TGF-beta R1 is targeted for lysosomal degradation thus maintaining inverse levels of these molecules. Finally, we show that E-CAD interacts with ERK, and that the decreased pluripotency upon CME loss can be rescued by inhibiting TGF-beta R, MEK, and GSK3 beta, or overexpressing E-CAD. Our results demonstrate that CME is critical for balancing signaling outputs to regulate ESC pluripotency, and possibly cell fate choices in early development.</p>1 a1 aGadgil, Chetan1 aMote, Ridim, D.1 aRajan, Raghav1 aSubramanyam, Deepa  uhttps://library.ncl.res.in/content/clathrin-mediated-endocytosis-regulates-balance-between-opposing-signals-maintain