Dengue Virus Nonstructural 3 Protein Interacts Directly with Human Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) and Reduces Its Glycolytic Activity

Overview

Journal Sci Rep

Specialty Science

Date 2019 Feb 27

PMID 30804377

Citations 17

Authors

Emiliana M Silva

Jonas N Conde

Diego Allonso

Gustavo T Ventura

Diego R Coelho

Pedro Henrique Carneiro

Manuela L Silva

Marciano V Paes

Kissila Rabelo

Gilberto Weissmuller

Paulo Mascarello Bisch

Ronaldo Mohana-Borges

Affiliations

Soon will be listed here.

Abstract

Dengue is an important mosquito-borne disease and a global public health problem. The disease is caused by dengue virus (DENV), which is a member of the Flaviviridae family and contains a positive single-stranded RNA genome that encodes a single precursor polyprotein that is further cleaved into structural and non-structural proteins. Among these proteins, the non-structural 3 (NS3) protein is very important because it forms a non-covalent complex with the NS2B cofactor, thereby forming the functional viral protease. NS3 also contains a C-terminal ATPase/helicase domain that is essential for RNA replication. Here, we identified 47 NS3-interacting partners using the yeast two-hybrid system. Among those partners, we highlight several proteins involved in host energy metabolism, such as apolipoprotein H, aldolase B, cytochrome C oxidase and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). GAPDH directly binds full-length NS3 and its isolated helicase and protease domains. Moreover, we observed an intense colocalization between the GAPDH and NS3 proteins in DENV2-infected Huh7.5.1 cells, in NS3-transfected BHK-21 cells and in hepatic tissue from a fatal dengue case. Taken together, these results suggest that the human GAPDH-DENV NS3 interaction is involved in hepatic metabolic alterations, which may contribute to the appearance of steatosis in dengue-infected patients. The interaction between GAPDH and full-length NS3 or its helicase domain in vitro as well as in NS3-transfected cells resulted in decreased GAPDH glycolytic activity. Reduced GAPDH glycolytic activity may lead to the accumulation of metabolic intermediates, shifting metabolism to alternative, non-glycolytic pathways. This report is the first to identify the interaction of the DENV2 NS3 protein with the GAPDH protein and to demonstrate that this interaction may play an important role in the molecular mechanism that triggers hepatic alterations.

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References

Sirover M . New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta. 1999; 1432(2):159-84. DOI: 10.1016/s0167-4838(99)00119-3. View

McGuffin L, Bryson K, Jones D . The PSIPRED protein structure prediction server. Bioinformatics. 2000; 16(4):404-5. DOI: 10.1093/bioinformatics/16.4.404. View

Tisdale E . Glyceraldehyde-3-phosphate dehydrogenase is required for vesicular transport in the early secretory pathway. J Biol Chem. 2000; 276(4):2480-6. DOI: 10.1074/jbc.M007567200. View

Dastoor Z, Dreyer J . Potential role of nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase in apoptosis and oxidative stress. J Cell Sci. 2001; 114(Pt 9):1643-53. DOI: 10.1242/jcs.114.9.1643. View

Leung D, Schroder K, White H, Fang N, Stoermer M, Abbenante G . Activity of recombinant dengue 2 virus NS3 protease in the presence of a truncated NS2B co-factor, small peptide substrates, and inhibitors. J Biol Chem. 2001; 276(49):45762-71. DOI: 10.1074/jbc.M107360200. View

Marti-Renom M, Madhusudhan M, Fiser A, Rost B, Sali A . Reliability of assessment of protein structure prediction methods. Structure. 2002; 10(3):435-40. DOI: 10.1016/s0969-2126(02)00731-1. View

Lamarre D, Anderson P, Bailey M, Beaulieu P, Bolger G, Bonneau P . An NS3 protease inhibitor with antiviral effects in humans infected with hepatitis C virus. Nature. 2003; 426(6963):186-9. DOI: 10.1038/nature02099. View

Comeau S, Gatchell D, Vajda S, Camacho C . ClusPro: an automated docking and discrimination method for the prediction of protein complexes. Bioinformatics. 2003; 20(1):45-50. DOI: 10.1093/bioinformatics/btg371. View

Chua J, Ng M, Chow V . The non-structural 3 (NS3) protein of dengue virus type 2 interacts with human nuclear receptor binding protein and is associated with alterations in membrane structure. Virus Res. 2004; 102(2):151-63. DOI: 10.1016/j.virusres.2004.01.025. View

10.

Comeau S, Gatchell D, Vajda S, Camacho C . ClusPro: a fully automated algorithm for protein-protein docking. Nucleic Acids Res. 2004; 32(Web Server issue):W96-9. PMC: 441492. DOI: 10.1093/nar/gkh354. View

11.

Bonafe N, Gilmore-Hebert M, Folk N, Azodi M, Zhou Y, Chambers S . Glyceraldehyde-3-phosphate dehydrogenase binds to the AU-Rich 3' untranslated region of colony-stimulating factor-1 (CSF-1) messenger RNA in human ovarian cancer cells: possible role in CSF-1 posttranscriptional regulation and tumor phenotype. Cancer Res. 2005; 65(9):3762-71. DOI: 10.1158/0008-5472.CAN-04-3954. View

12.

Hara M, Snyder S . Nitric oxide-GAPDH-Siah: a novel cell death cascade. Cell Mol Neurobiol. 2006; 26(4-6):527-38. PMC: 11520605. DOI: 10.1007/s10571-006-9011-6. View

13.

Kozakov D, Brenke R, Comeau S, Vajda S . PIPER: an FFT-based protein docking program with pairwise potentials. Proteins. 2006; 65(2):392-406. DOI: 10.1002/prot.21117. View

14.

Freire M, Marchevsky R, Almeida L, Yamamura A, Caride E, Brindeiro P . Wild dengue virus types 1, 2 and 3 viremia in rhesus monkeys. Mem Inst Oswaldo Cruz. 2007; 102(2):203-8. DOI: 10.1590/s0074-02762007005000011. View

15.

Negi S, Schein C, Oezguen N, Power T, Braun W . InterProSurf: a web server for predicting interacting sites on protein surfaces. Bioinformatics. 2007; 23(24):3397-9. PMC: 2636624. DOI: 10.1093/bioinformatics/btm474. View

16.

Luo D, Xu T, Hunke C, Gruber G, Vasudevan S, Lescar J . Crystal structure of the NS3 protease-helicase from dengue virus. J Virol. 2007; 82(1):173-83. PMC: 2224403. DOI: 10.1128/JVI.01788-07. View

17.

El-Bacha T, Midlej V, Da Silva A, Silva da Costa L, Benchimol M, Galina A . Mitochondrial and bioenergetic dysfunction in human hepatic cells infected with dengue 2 virus. Biochim Biophys Acta. 2007; 1772(10):1158-66. DOI: 10.1016/j.bbadis.2007.08.003. View

18.

Lee D, Redfern O, Orengo C . Predicting protein function from sequence and structure. Nat Rev Mol Cell Biol. 2007; 8(12):995-1005. DOI: 10.1038/nrm2281. View

19.

Bente D, Rico-Hesse R . Models of dengue virus infection. Drug Discov Today Dis Models. 2007; 3(1):97-103. PMC: 1949394. DOI: 10.1016/j.ddmod.2006.03.014. View

20.

Eswar N, Eramian D, Webb B, Shen M, Sali A . Protein structure modeling with MODELLER. Methods Mol Biol. 2008; 426:145-59. DOI: 10.1007/978-1-60327-058-8_8. View