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Glutamine deprivation initiates reversible assembly of mammalian rods and rings

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Abstract

Rods and rings (RR) are protein assemblies composed of cytidine triphosphate synthetase type 1 (CTPS1) and inosine monophosphate dehydrogenase type 2 (IMPDH2), key enzymes in CTP and GTP biosynthesis. Small-molecule inhibitors of CTPS1 or IMPDH2 induce RR assembly in various cancer cell lines within 15 min to hours. Since glutamine is an essential amide nitrogen donor in these nucleotide biosynthetic pathways, glutamine deprivation was examined to determine whether it leads to RR formation. HeLa cells cultured in normal conditions did not show RR, but after culturing in media lacking glutamine, short rods (<2 μm) assembled after 24 h, and longer rods (>5 μm) formed after 48 h. Upon supplementation with glutamine or guanosine, these RR underwent almost complete disassembly within 15 min. Inhibition of glutamine synthetase with methionine sulfoximine also increased RR assembly in cells deprived of glutamine. Taken together, our data support the hypothesis that CTP/GTP biosynthetic enzymes polymerize to form RR in response to a decreased intracellular level of glutamine. We speculate that rod and ring formation is an adaptive metabolic response linked to disruption of glutamine homeostasis.

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Abbreviations

RR:

Rods and rings

CTPS:

Cytidine triphosphate synthetase

IMPDH:

Inosine monophosphate dehydrogenase

DON:

6-Diazo-5-oxo-l-norleucine

CTP:

Cytidine triphosphate

GMP:

Guanine monophosphate

MPA:

Mycophenolic acid

FBS:

Fetal bovine serum

GTP:

Guanosine triphosphate

UTP:

Uridine triphosphate

mRNA:

Messenger RNA

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Acknowledgments

We thank Thuy Nguyen and Dania Saleem for their technical assistance with ribonucleoside treatment and time-point analysis of RR disassembly by glutamine.

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Authors and Affiliations

  1. Department of Oral Biology, University of Florida, 1395 Center Drive, Gainesville, FL, 32610-0424, USA

    S. John Calise, Wendy C. Carcamo, Claire Krueger, Joyce D. Yin & Edward K. L. Chan

  2. Department of Biochemistry and Molecular Biology, University of Florida, 1600 SW Archer Rd., Gainesville, FL, 32610-0245, USA

    Daniel L. Purich

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  1. S. John Calise
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  2. Wendy C. Carcamo
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  3. Claire Krueger
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Correspondence to Edward K. L. Chan.

Additional information

S. J. Calise and W. C. Carcamo contributed equally to this work.

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Calise, S.J., Carcamo, W.C., Krueger, C. et al. Glutamine deprivation initiates reversible assembly of mammalian rods and rings. Cell. Mol. Life Sci. 71, 2963–2973 (2014). https://doi.org/10.1007/s00018-014-1567-6

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  • DOI: https://doi.org/10.1007/s00018-014-1567-6

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