Only One Isoform of CTP Synthase Forms the Cytoophidium

English version České info

CTP synthase is an essential enzyme that plays a key role in energy metabolism. Several independent studies have demonstrated that CTP synthase can form an evolutionarily conserved subcellular structure termed cytoophidium. In budding yeast, there are two isoforms of CTP synthase and both isoforms localize in cytoophidium. However, little is known about the distribution of CTP synthase isoforms in Drosophila melanogaster. Here, we report that three transcripts generated at the CTP synthase gene locus exhibit different expression profiles, and three isoforms encoded by this gene locus show a distinct subcellular distribution. While isoform A localizes in the nucleus, isoform B distributes diffusely in the cytoplasm, and only isoform C forms the cytoophidium. In the two isoform C-specific mutants, cytoophidia disappear in the germline cells. Although isoform A does not localize to the cytoophidium, a mutation disrupting mostly isoform A expression results in the disassembly of cytoophidia. Overexpression of isoform C can induce the growth of the cytoophidium in a cell-autonomous manner. Ectopic expression of the cytoophidium-forming isoform does not cause any defect in the embryos. In addition, we identify that a small segment at the amino terminus of isoform C is necessary but not sufficient for cytoophidium formation. Finally, we demonstrate that an excess of the synthetase domain of CTP synthase disrupts cytoophidium formation. Thus, the study of multiple isoforms of CTP synthase in Drosophila provides a good opportunity to dissect the biogenesis and function of the cytoophidum in a genetically tractable organism.

Vyšlo v časopise: Only One Isoform of CTP Synthase Forms the Cytoophidium. PLoS Genet 9(2): e32767. doi:10.1371/journal.pgen.1003256
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003256

Souhrn

Zdroje

1. Alberts B, Johnson A, Lewis J, Raff M, Roberts K, et al.. (2008) Molecular Biology of the Cell, 5th Edition. New York/Abingdon: Garland Science, Taylor & Francis Group.

2. Nover L, Lynen F, Mothes K, editors (1980) Cell Compartmentation and Metabolic Channeling. Amsterdam -⁠ New York -⁠ Oxford: Elsevier North-Holland Inc. 523 p.

3. RazaH (2011) Dual localization of glutathione S-transferase in the cytosol and mitochondria: implications in oxidative stress, toxicity and disease. FEBS J 278 : 4243–4251.

4. NunnariJ, SuomalainenA (2012) Mitochondria: in sickness and in health. Cell 148 : 1145–1159.

5. BoyerPD (1997) The ATP synthase–a splendid molecular machine. Annu Rev Biochem 66 : 717–749.

6. LiebermanI (1956) Enzymatic amination of uridine triphosphate to cytidine triphosphate. J Biol Chem 222 : 765–775.

7. KammenHO, HurlbertRB (1959) The formation of cytidine nucleotides and RNA cytosine from orotic acid by the Novikoff tumor in vitro. Cancer Res 19 : 654–663.

8. ChakrabortyKP, HurlbertRB (1961) Role of glutamine in the biosynthesis of cytidine nucleotides in Escherichia coli. Biochim Biophys Acta 47 : 607–609.

9. LongCW, PardeeAB (1967) Cytidine triphosphate synthetase of Escherichia coli B. I. Purification and kinetics. J Biol Chem 242 : 4715–4721.

10. LevitzkiA, KoshlandDEJr (1971) Cytidine triphosphate synthetase. Covalent intermediates and mechanisms of action. Biochemistry 10 : 3365–3371.

11. von der SaalW, AndersonPM, VillafrancaJJ (1985) Mechanistic investigations of Escherichia coli cytidine-5′-triphosphate synthetase. Detection of an intermediate by positional isotope exchange experiments. J Biol Chem 260 : 14993–14997.

12. LewisDA, VillafrancaJJ (1989) Investigation of the mechanism of CTP synthetase using rapid quench and isotope partitioning methods. Biochemistry 28 : 8454–8459.

13. EndrizziJA, KimH, AndersonPM, BaldwinEP (2004) Crystal structure of Escherichia coli cytidine triphosphate synthetase, a nucleotide-regulated glutamine amidotransferase/ATP-dependent amidoligase fusion protein and homologue of anticancer and antiparasitic drug targets. Biochemistry 43 : 6447–6463.

14. EndrizziJA, KimH, AndersonPM, BaldwinEP (2005) Mechanisms of product feedback regulation and drug resistance in cytidine triphosphate synthetases from the structure of a CTP-inhibited complex. Biochemistry 44 : 13491–13499.

15. WilliamsJC, KizakiH, WeberG, MorrisHP (1978) Increased CTP synthetase activity in cancer cells. Nature 271 : 71–73.

16. KizakiH, WilliamsJC, MorrisHP, WeberG (1980) Increased cytidine 5′-triphosphate synthetase activity in rat and human tumors. Cancer Res 40 : 3921–3927.

17. VerschuurAC, van GennipAH, MullerEJ, VoutePA, van KuilenburgAB (1998) Increased activity of cytidine Triphosphate synthetase in pediatric acute lymphoblastic leukemia. Adv Exp Med Biol 431 : 667–671.

18. WhelanJ, SmithT, PhearG, RohatinerA, ListerA, et al. (1994) Resistance to cytosine arabinoside in acute leukemia: the significance of mutations in CTP synthetase. Leukemia 8 : 264–265.

19. van den BergAA, van LentheH, BuschS, de KorteD, RoosD, et al. (1993) Evidence for transformation-related increase in CTP synthetase activity in situ in human lymphoblastic leukemia. Eur J Biochem 216 : 161–167.

20. van den BergAA, van LentheH, KippJB, de KorteD, van KuilenburgAB, et al. (1995) Cytidine triphosphate (CTP) synthetase activity during cell cycle progression in normal and malignant T-lymphocytic cells. Eur J Cancer 31A: 108–112.

21. WeberG, LuiMS, TakedaE, DentonJE (1980) Enzymology of human colon tumors. Life Sci 27 : 793–799.

22. VerschuurAC, van GennipAH, BrinkmanJ, VoutePA, van KuilenburgAB (2000) Cyclopentenyl cytosine induces apoptosis and secondary necrosis in a T-lymphoblastic leukemic cell-line. Adv Exp Med Biol 486 : 319–325.

23. VerschuurAC, van GennipAH, LeenR, VoutePA, van KuilenburgAB (2000) Cyclopentenyl cytosine increases the phosphorylation and incorporation into dna of arabinofu-ranosyl cytosine in a myeloid leukemic cell-line. Adv Exp Med Biol 486 : 311–317.

24. VerschuurAC, Van GennipAH, LeenR, MeinsmaR, VoutePA, et al. (2000) In vitro inhibition of cytidine triphosphate synthetase activity by cyclopentenyl cytosine in paediatric acute lymphocytic leukaemia. Br J Haematol 110 : 161–169.

25. VerschuurAC, Van GennipAH, LeenR, MullerEJ, ElzingaL, et al. (2000) Cyclopentenyl cytosine inhibits cytidine triphosphate synthetase in paediatric acute non-lymphocytic leukaemia: a promising target for chemotherapy. Eur J Cancer 36 : 627–635.

26. VerschuurAC, BrinkmanJ, Van GennipAH, LeenR, VetRJ, et al. (2001) Cyclopentenyl cytosine induces apoptosis and increases cytarabine-induced apoptosis in a T-lymphoblastic leukemic cell-line. Leuk Res 25 : 891–900.

27. EllimsPH, GanTE, MedleyG (1983) Cytidine triphosphate synthetase activity in lymphoproliferative disorders. Cancer Res 43 : 1432–1435.

28. De ClercqE (2001) Vaccinia virus inhibitors as a paradigm for the chemotherapy of poxvirus infections. Clin Microbiol Rev 14 : 382–397.

29. FijolekA, HoferA, ThelanderL (2007) Expression, purification, characterization, and in vivo targeting of trypanosome CTP synthetase for treatment of African sleeping sickness. J Biol Chem 282 : 11858–11865.

30. HoferA, SteverdingD, ChabesA, BrunR, ThelanderL (2001) Trypanosoma brucei CTP synthetase: a target for the treatment of African sleeping sickness. Proc Natl Acad Sci U S A 98 : 6412–6416.

31. HendriksEF, O'SullivanWJ, StewartTS (1998) Molecular cloning and characterization of the Plasmodium falciparum cytidine triphosphate synthetase gene. Biochim Biophys Acta 1399 : 213–218.

32. WylieJL, BerryJD, McClartyG (1996) Chlamydia trachomatis CTP synthetase: molecular characterization and developmental regulation of expression. Mol Microbiol 22 : 631–642.

33. LiuJL (2010) Intracellular compartmentation of CTP synthase in Drosophila. J Genet Genomics 37 : 281–296.

34. Ingerson-MaharM, BriegelA, WernerJN, JensenGJ, GitaiZ (2010) The metabolic enzyme CTP synthase forms cytoskeletal filaments. Nat Cell Biol 12 : 739–746.

35. NoreeC, SatoBK, BroyerRM, WilhelmJE (2010) Identification of novel filament-forming proteins in Saccharomyces cerevisiae and Drosophila melanogaster. J Cell Biol 190 : 541–551.

36. ChenK, ZhangJ, TastanOY, DeussenZA, SiswickMY, et al. (2011) Glutamine analogs promote cytoophidium assembly in human and Drosophila cells. J Genet Genomics 38 : 391–402.

37. CarcamoWC, SatohM, KasaharaH, TeradaN, HamazakiT, et al. (2011) Induction of cytoplasmic rods and rings structures by inhibition of the CTP and GTP synthetic pathway in mammalian cells. PLoS ONE 6: e29690 doi:10.1371/journal.pone.0029690.

38. LiuJL (2011) The enigmatic cytoophidium: compartmentation of CTP synthase via filament formation. Bioessays 33 : 159–164.

39. GillilandWD, ViettiDL, SchweppeNM, GuoF, JohnsonTJ, et al. (2009) Hypoxia transiently sequesters mps1 and polo to collagenase-sensitive filaments in Drosophila prometaphase oocytes. PLoS ONE 4: e7544 doi:10.1371/journal.pone.0007544.

40. BhaskarV, CoureyAJ (2002) The MADF-BESS domain factor Dip3 potentiates synergistic activation by Dorsal and Twist. Gene 299 : 173–184.

41. BuszczakM, PaternoS, LighthouseD, BachmanJ, PlanckJ, et al. (2007) The carnegie protein trap library: a versatile tool for Drosophila developmental studies. Genetics 175 : 1505–1531.

42. CraymerL (1980) [New mutants report.]. Drosophila Information Service 55 : 197–200.

43. Spradling AC (1993) Developmental genetics of oogenesis. In: Bate M, Martinez Arias A, editors. The Development of Drosophila melanogaster: Cold Spring Harbor Laboratory Press. pp. 1–70.

44. RobertsonJG (1995) Determination of subunit dissociation constants in native and inactivated CTP synthetase by sedimentation equilibrium. Biochemistry 34 : 7533–7541.

45. ZalkinH (1993) The amidotransferases. Adv Enzymol Relat Areas Mol Biol 66 : 203–309.

46. ZalkinH, SmithJL (1998) Enzymes utilizing glutamine as an amide donor. Adv Enzymol Relat Areas Mol Biol 72 : 87–144.

47. MassiereF, Badet-DenisotMA (1998) The mechanism of glutamine-dependent amidotransferases. Cell Mol Life Sci 54 : 205–222.

48. WengML, ZalkinH (1987) Structural role for a conserved region in the CTP synthetase glutamine amide transfer domain. J Bacteriol 169 : 3023–3028.

49. LevitzkiA, KoshlandDEJr (1976) The role of negative cooperativity and half-of-the-sites reactivity in enzyme regulation. Curr Top Cell Regul 10 : 1–40.

50. KozhevnikovaEN, van der KnaapJA, PindyurinAV, OzgurZ, van IjckenWF, et al. (2012) Metabolic enzyme IMPDH is also a transcription factor regulated by cellular state. Mol Cell 47 : 133–139.

51. ZeccaM, StruhlG (2002) Subdivision of the Drosophila wing imaginal disc by EGFR-mediated signaling. Development 129 : 1357–1368.

52. ThibaultST, SingerMA, MiyazakiWY, MilashB, DompeNA, et al. (2004) A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac. Nat Genet 36 : 283–287.

53. NagaiT, IbataK, ParkES, KubotaM, MikoshibaK, et al. (2002) A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat Biotechnol 20 : 87–90.

54. ThomsenS, AzzamG, KaschulaR, WilliamsLS, AlonsoCR (2010) Developmental RNA processing of 3′UTRs in Hox mRNAs as a context-dependent mechanism modulating visibility to microRNAs. Development 137 : 2951–2960.

55. BischofJ, MaedaRK, HedigerM, KarchF, BaslerK (2007) An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases. Proc Natl Acad Sci U S A 104 : 3312–3317.