Transcriptomic analysis of differential gene expression reveals an increase in COX2 levels during <i>in vitro</i> canine herpesvirus infection

Kurissio, Jacqueline Kazue; Araújo Júnior, João Pessoa

doi:10.1590/0103-8478cr20170945

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MICROBIOLOGY • Cienc. Rural 48 (10) • 2018 • https://doi.org/10.1590/0103-8478cr20170945 copy

Transcriptomic analysis of differential gene expression reveals an increase in COX2 levels during in vitro canine herpesvirus infection

Análise transcriptômica da expressão gênica diferencial revela um aumento nos níveis de COX2 durante a infecção in vitro por herpesvirus canino

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT:

Canine herpesvirus (CaHV-1) affects canids worldwide, causing death in neonates and immunosuppressed hosts. Acute infection by CaHV-1 can cause reproductive, respiratory, and neurological problems in adult animals. Viral pathogenesis and host genes expressions during of CaHV-1infection are not clearly understood. In the present study, the transcriptome of canine kidney cell Mardin-Darby (MDCK) infected in vitro with canine herpesvirus was explored. For this, RNA sequencing (RNA-seq) of the samples in different moments during infection was carried out. Subsequently, the transcriptomic analysis genes related to cell activities and process involved to viral cycle infection were evaluated until 32h post-inoculation (pi). Among evaluated genes, was verified a significant and gradative increase of the prostaglandin-endoperoxide synthase 2 (PTGS2) or cyclooxygenase 2 (COX2) gene expression, throughout of infection, though differential gene expression analysis and validated by quantitative reverse transcription PCR (RT-qPCR). High COX2 expression is usually induced in response to inflammation, pathogens or activation of the immune system but can be a viral mechanism to favor viral replication. Thus, COX2 level increase can be a favorable factor for viral infection with Cahv-1 virus and the use of selective COX2 inhibitors may be beneficial for limiting the infection or clinical signs by causing interruption of the viral replication cycle during active infection. Additionally, the regulation genes expression differential verified in this study can contribute to determining important targets for inhibiting canine herpesvirus infection either by cellular or viral mechanisms.

Key words:
transcriptome; herpesvirus; CaHV-1; PTGS2; COX2; canine pathogen

Universidade Federal de Santa Maria Universidade Federal de Santa Maria, Centro de Ciências Rurais , 97105-900 Santa Maria RS Brazil , Tel.: +55 55 3220-8698 , Fax: +55 55 3220-8695 - Santa Maria - RS - Brazil
E-mail: cienciarural@mail.ufsm.br

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[1] E-mail: jkkurissio@yahoo.com.br. *Corresponding author

Gene Symbol	Description	Gene product function	Time	FC
1. EGR-1	Early growth response protein 1	Transcriptional regulator	1h	35.30
2. FOS	Proto-oncogene c-Fos	Essential for cell proliferation and differentiation	1h	33.38
3. COX2	Prostaglandin g/h synthase 2	Increase cell adhesion, phenotypic changes, resistance to apoptosis and tumor angiogenesis	32h	26.41
4. CHAC1	Cha C glutathione specific gamma-glutamylcyclotransferase 1	Pro-apoptotic component	32h	24.74
5. DDIT4	DNA damage-inducible transcript 4 protein	Responses to cellular energy levels and cellular stress, including responses to hypoxia and DNA damage.	32h	20.83
6. ATF3	Cyclic amp-dependent transcription factor ATF-3	This protein binds the camp response element (CRE), present in many viral and cellular promoters, repressor of inflammatory response	32h	20.37
7. ISG20	Interferon stimulated exonuclease gene 20	Interferon-induced antiviral exoribonuclease	32h	12.75
8. BTG2	BTG anti-proliferation factor2	Anti-proliferative protein	32h	11.44
9. RPL4	RPL4 ribosomalprotein L4	Compose a large ribonucleoprotein complex responsible for the synthesis of proteins in the cell	1h	11.13
10. DUSP1	Dual specificity protein phosphatase 1	Cellular response to environmental stress	1h	10.79
11. MARCKSL1	MARCKS like1	Controls cell movement by regulating actin cytoskeleton homeostasis	32h	9.79
12. TNFAIP3	TNF alpha inducedprotein3	Involved in the cytokine-mediated immune and inflammatory responses.	32h	9.52
13. SON	Protein SON	DNA-binding, tumorigenesis and apoptosis, but functional significance of this protein had not been recognized.	16h	9.42
14. MRPL33_1	Mitochondrial ribosomal protein L33	Large mitoribosomal subunit protein, possible involvement in mitochondrial translation.	16h	9.14
15. YPEL3	Yippee like 3	Inhibitory effects in normal and tumor cell lines	32h	9.08
16. SMIM10L1	Small integral membrane protein 10 like 1	The function SMIM10L1 is not clear	16h	8.13
17. MYH14	Myosin heavy chain 14	Regulate the cytokinesis, motility and polarity cell	28h	7.76
18. EIF4A1	Eukaryotic translation initiation factor 4A1	Proliferation and growth cell	16h	7.38
19. RPL37	Ribosomal protein L37	Component of the 60S subunit	16h	6.61
20. UBALD1	UBA like domain containing 1	No characterized	0h	6.6

Gene Symbol	Description	Encodes function protein	Time	FC
1. RPL26	Ribosomal protein L26	Cell proliferation	24h	-236.25
2. ATP5G2	ATP synthase, H+ transporting, mitochondrial F0 complex, subunitc-2	Mitochondrial ATP synthesis coupled proton transport	16h	-99.11
3. ATP5MC2	ATP synthase membrane subunit c locus 2	Lipid binding and proton transmembrane transporter activity	4h	-46.36
4. TMED9	Trans membrane p24 trafficking protein 9	Transport proteins located in the endoplasmic reticulum and Golgi complex	16h	-38.05
5. RPL23A	Ribosomal protein L23a	Cell growth and regulation	0h	-35.08
6. RPS27	Ribosomal protein S27 or metallopan-stimulin 1 (MPS-1)	Extra-ribosomal functions, regulate growth and survival cell as well as DNA repair and transcription	0h	-26.26
7. ARPC3	Actin related protein 2/3 complex subunit 3	Control of actin polymerization in cells	4h	-24.84
8. RPL29	RPL29 ribosomal protein L29	Cell proliferation	16h	-24.54
9. S100A2	S100 calcium binding protein A2	Cell regulation	16h	-19.79
10. CRYAB	Crystallin alpha B	Apoptosis inhibition	0h	-15.40
11. EEF1A1	Eukaryotice longation factor 1 alpha	Instead to upregulation, the down regulation of eEF1A1, is required for the apoptosis	4h	-14.12
12. TMEM97	Transmembrane protein 97	Cell proliferation	16h	-12.94
13. DLADRA	MHC class II DR alpha chain	Cellular immunity	28h	-12.26
14. MRPS17	Mitochondrial ribosomal protein S17	Mitochondrial proteins synthesis	4h	-11.17
15. DHRS2	Dehydrogenase/reductase2	Cycle cell regulation and apoptosis	28h	-10.95
16. MMP13	Matrix metallopeptidase 13	Cell proliferation	4h	-10.68
17. KRT14	Keratin 14	Structural molecule activity and keratin filament binding	0h	-10.65
18. SMIM7	Small integral membrane protein 7	Integral component membrane	4h	-10.39
19. TMEM179B	Transmembrane protein 179B	Membrane protein transporter	0h	-9.71
20. CCL17	C-C motif chemokine ligand 17	Mediating by chemotaxis the infiltration of Th2 cells in inflammatory reaction	4h	-9.69