Fic-mediated DeAMPylation is Not Dependent on Homodimerization and Rescues Toxic AMPylation in Flies

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2017 Nov 2

PMID 29089387

Citations 26

Authors

Amanda K Casey

Andrew T Moehlman

Junmei Zhang

Kelly A Servage

Helmut Kramer

Kim Orth

Affiliations

Soon will be listed here.

Abstract

Protein chaperones play a critical role in proteostasis. The activity of the major endoplasmic reticulum chaperone BiP (GRP78) is regulated by Fic-mediated AMPylation during resting states. By contrast, during times of stress, BiP is deAMPylated. Here, we show that excessive AMPylation by a constitutively active Fic mutant is lethal in This lethality is cell-autonomous, as directed expression of the mutant Fic to the fly eye does not kill the fly but rather results in a rough and reduced eye. Lethality and eye phenotypes are rescued by the deAMPylation activity of wild-type Fic. Consistent with Fic acting as a deAMPylation enzyme, its activity was both time- and concentration-dependent. Furthermore, Fic deAMPylation activity was sufficient to suppress the AMPylation activity mediated by the constitutively active Fic mutant in S2 lysates. Further, we show that the dual enzymatic activity of Fic is, in part, regulated by Fic dimerization, as loss of this dimerization increases AMPylation and reduces deAMPylation of BiP.

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References

Vasudevan D, Ryoo H . Detection of Cell Death in Drosophila Tissues. Methods Mol Biol. 2016; 1419:131-44. PMC: 4867113. DOI: 10.1007/978-1-4939-3581-9_11. View

Sanyal A, Chen A, Nakayasu E, Lazar C, Zbornik E, Worby C . A novel link between Fic (filamentation induced by cAMP)-mediated adenylylation/AMPylation and the unfolded protein response. J Biol Chem. 2015; 290(13):8482-99. PMC: 4375499. DOI: 10.1074/jbc.M114.618348. View

Yarbrough M, Li Y, Kinch L, Grishin N, Ball H, Orth K . AMPylation of Rho GTPases by Vibrio VopS disrupts effector binding and downstream signaling. Science. 2008; 323(5911):269-72. DOI: 10.1126/science.1166382. View

Ham H, Woolery A, Tracy C, Stenesen D, Kramer H, Orth K . Unfolded protein response-regulated Drosophila Fic (dFic) protein reversibly AMPylates BiP chaperone during endoplasmic reticulum homeostasis. J Biol Chem. 2014; 289(52):36059-69. PMC: 4276871. DOI: 10.1074/jbc.M114.612515. View

Kramer H, Phistry M . Mutations in the Drosophila hook gene inhibit endocytosis of the boss transmembrane ligand into multivesicular bodies. J Cell Biol. 1996; 133(6):1205-15. PMC: 2120908. DOI: 10.1083/jcb.133.6.1205. View

Rahman M, Ham H, Liu X, Sugiura Y, Orth K, Kramer H . Visual neurotransmission in Drosophila requires expression of Fic in glial capitate projections. Nat Neurosci. 2012; 15(6):871-5. PMC: 3578554. DOI: 10.1038/nn.3102. View

Nandi N, Tyra L, Stenesen D, Kramer H . Acinus integrates AKT1 and subapoptotic caspase activities to regulate basal autophagy. J Cell Biol. 2014; 207(2):253-68. PMC: 4210446. DOI: 10.1083/jcb.201404028. View

Preissler S, Chambers J, Crespillo-Casado A, Avezov E, Miranda E, Perez J . Physiological modulation of BiP activity by trans-protomer engagement of the interdomain linker. Elife. 2015; 4:e08961. PMC: 4608358. DOI: 10.7554/eLife.08961. View

Garcia-Pino A, Zenkin N, Loris R . The many faces of Fic: structural and functional aspects of Fic enzymes. Trends Biochem Sci. 2014; 39(3):121-9. DOI: 10.1016/j.tibs.2014.01.001. View

10.

Brown M, Segal A, Stadtman E . Modulation of glutamine synthetase adenylylation and deadenylylation is mediated by metabolic transformation of the P II -regulatory protein. Proc Natl Acad Sci U S A. 1971; 68(12):2949-53. PMC: 389567. DOI: 10.1073/pnas.68.12.2949. View

11.

Kingdon H, SHAPIRO B, Stadtman E . Regulation of glutamine synthetase. 8. ATP: glutamine synthetase adenylyltransferase, an enzyme that catalyzes alterations in the regulatory properties of glutamine synthetase. Proc Natl Acad Sci U S A. 1967; 58(4):1703-10. PMC: 223983. DOI: 10.1073/pnas.58.4.1703. View

12.

Ray M, Lakhotia S . The commonly used eye-specific sev-GAL4 and GMR-GAL4 drivers in Drosophila melanogaster are expressed in tissues other than eyes also. J Genet. 2015; 94(3):407-16. DOI: 10.1007/s12041-015-0535-8. View

13.

Truttmann M, Zheng X, Hanke L, Damon J, Grootveld M, Krakowiak J . Unrestrained AMPylation targets cytosolic chaperones and activates the heat shock response. Proc Natl Acad Sci U S A. 2016; 114(2):E152-E160. PMC: 5240723. DOI: 10.1073/pnas.1619234114. View

14.

Gratz S, Cummings A, Nguyen J, Hamm D, Donohue L, Harrison M . Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Genetics. 2013; 194(4):1029-35. PMC: 3730909. DOI: 10.1534/genetics.113.152710. View

15.

Hao Y, Chuang T, Ball H, Luong P, Li Y, Flores-Saaib R . Characterization of a rabbit polyclonal antibody against threonine-AMPylation. J Biotechnol. 2010; 151(3):251-4. PMC: 4391625. DOI: 10.1016/j.jbiotec.2010.12.013. View

16.

Venken K, He Y, Hoskins R, Bellen H . P[acman]: a BAC transgenic platform for targeted insertion of large DNA fragments in D. melanogaster. Science. 2006; 314(5806):1747-51. DOI: 10.1126/science.1134426. View

17.

Ceriani M . Basic protocols for Drosophila S2 cell line: maintenance and transfection. Methods Mol Biol. 2007; 362:415-22. DOI: 10.1007/978-1-59745-257-1_33. View

18.

Wolff T . Preparation of Drosophila eye specimens for scanning electron microscopy. Cold Spring Harb Protoc. 2011; 2011(11):1383-5. DOI: 10.1101/pdb.prot066506. View

19.

Kozutsumi Y, Segal M, Normington K, Gething M, Sambrook J . The presence of malfolded proteins in the endoplasmic reticulum signals the induction of glucose-regulated proteins. Nature. 1988; 332(6163):462-4. DOI: 10.1038/332462a0. View

20.

Truttmann M, Cruz V, Guo X, Engert C, Schwartz T, Ploegh H . The Caenorhabditis elegans Protein FIC-1 Is an AMPylase That Covalently Modifies Heat-Shock 70 Family Proteins, Translation Elongation Factors and Histones. PLoS Genet. 2016; 12(5):e1006023. PMC: 4854385. DOI: 10.1371/journal.pgen.1006023. View