Cloning and differential expression analyses of Cdc42 from sheep

Open access

Abstract

Introduction: Serological diagnosis of brucellosis is still a great challenge due to the infeasibility of discriminating infected animals from vaccinated ones, so it is necessary to search for diagnostic biomarkers for differential diagnosis of brucellosis.

Material and Methods: Cell division cycle 42 (Cdc42) from sheep (Ovis aries) (OaCdc42) was cloned by rapid amplification of cDNA ends (RACE), and then tissue distribution and differential expression levels of OaCdc42 mRNA between infected and vaccinated sheep were analysed by RT-qPCR.

Results: The full-length cDNA of OaCdc42 was 1,609 bp containing an open reading frame (ORF) of 576 bp. OaCdc42 mRNAs were detected in the heart, liver, spleen, lung, kidneys, rumen, small intestine, skeletal muscles, and buffy coat, and the highest expression was detected in the small intestine. Compared to the control, the levels of OaCdc42 mRNA from sheep infected with Brucella melitensis or sheep vaccinated with Brucella suis S2 was significantly different (P < 0.01) after 40 and 30 days post-inoculation, respectively. However, the expression of OaCdc42 mRNA was significantly different between vaccinated and infected sheep (P < 0.05 or P < 0.01) on days: 14, 30, and 60 post-inoculation, whereas no significant difference (P > 0.05) was noted 40 days post-inoculation. Moreover, the expression of OaCdc42 from both infected and vaccinated sheep showed irregularity.

Conclusion: OaCdc42 is not a good potential diagnostic biomarker for differential diagnosis of brucellosis in sheep.

1. Ahmed W., Zheng K., Liu Z.F.: Establishment of chronic infection: Brucella's Stealth Strategy. Frontiers Cell Infect Microbiol 2016, 6, 1–12.

2. Deevi R., Fatehullah A., Jagan I., Nagaraju M., Bingham V., Campbell F.C.: PTEN regulates colorectal epithelial apoptosis through Cdc42 signalling. Br J Cancer 2011, 105, 1313–1321.

3. Dorneles E.M.S., Teixeira-Carvalho A., Araujo M.S.S., Sriranganathan N., Lage A.P.: Immune response triggered by Brucella abortus following infection or vaccination. Vaccine 2015, 33, 3659–3666.

4. Farhan H., Hsu V.W.: Cdc42 and cellular polarity: emercing roles at the Golgi. Trends Cell Biol 2016, 26, 241–248.

5. Franco M.P., Mulder M., Gilman R.H., Smits H.L.: Human brucellosis. Lancet Infect Dis 2007, 7, 775–786.

6. Guzman-Verri C., Chaves-Olarte E., von Eichel-Streiber C., Lopez-Goni I., Thelestam M., Arvidson S., Gorvel J.P., Moreno E.: GTPases of the Rho subfamily are required for Brucella abortus internalization in nonprofessional phagocytes - direct activation of Cdc42. J Biol Chem 2001, 276, 44435–44443.

7. Hao Y., Zhang X., Zhang D., Cheng Y., Du L., Kuang W., Lei M., Jiao H., Qi C., Wang F.: Molecular cloning and tissue expression of cell division cycle 42 in buffalo tissue. J Anim Vet Adv 2012, 11, 1018–1022.

8. Hu J.H., Mukhopadhyay A., Craig A.W.B.: Transducer of Cdc42-dependent actin assembly promotes epidermal growth factor-induced cell motility and invasiveness. J Biol Chem 2011, 286, 2261–2272.

9. Keestra A.M., Winter M.G., Auburger J.J., Fraessle S.P., Xavier M.N., Winter S.E., Kim A., Poon V., Ravesloot M.M., Waldenmaier J.F.T., Tsolis R.M., Eigenheer R.A., Bauemler A.J.: Manipulation of small Rho GTPases is a pathogen-induced process detected by NOD1. Nature 2013, 496, 233–237.

10. Lee K., Boyd K.L., Parekh D.V., Kehl-Fie T.E., Baldwin H.S., Brakebusch C., Skaar E.P., Boothby M., Zent R.: Cdc42 promotes host defenses against fatal infection. Infect Immun 2013, 81, 2714–2723.

11. Liu G.Y., Gao S.Z., Ge C.R., Zhang X.: cDNA cloning and tissue expression analyses of the encoding regions for three novel porcine genes-MJD1, CDC42, and NECD. Anim Biotechnol 2008, 19, 117–121.

12. Melendez J., Liu M., Sampson L., Akunuru S., Han X., Vallance J., Witte D., Shroyer N., Zheng Y.: Cdc42 coordinates proliferation, polarity, migration, and differentiation of small intestinal epithelial cells in mice. Gastroenterology 2013, 145, 808–819.

13. Nielsen K., Smith P., Widdison J., Gall D., Kelly L., Kelly W., Nicoletti P.: Serological relationship between cattle exposed to Brucella abortus, Yersinia enterocolitica O:9, and Escherichia coli O157:H7. Vet Microbiol 2004, 100, 25–30.

14. Patel J.C., Galan J.E.: Differential activation and function of Rho GTPases during Salmonella-host cell interactions. J Cell Biol 2006, 175, 453–463.

15. Raftopoulou M., Hall A.: Cell migration: Rho GTPases lead the way. Dev Biol 2004, 265, 23–32.

16. Rigano L.A., Dowd G.C., Wang Y., Ireton K.: Listeria monocytogenes antagonizes the human GTPase Cdc42 to promote bacterial spread. Cell Microbiol 2014, 16, 1068–1079.

17. Swaine T., Dittmar M.T.: CDC42 use in viral cell entry processes by RNA viruses. Viruses-Basel 2015, 7, 6526–6536.

18. Wang J.-Y., Wu N., Liu W.-H., Ren J.-J., Tang P., Qiu Y.-H., Wang C.-Y., Chang C.-D., Liu H.-J.: A repA-based ELISA for discriminating cattle vaccinated with Brucella suis 2 from those naturally infected with Brucella abortus and Brucella melitensis. Mol Cell Probes 2014, 28, 251–254.

19. Watson L.J., Rossi G., Brennwald P.: Quantitative analysis of membrane trafficking in regulation of Cdc42 polarity. Traffic 2014, 15, 1330–1343.

20. Xu J.-D., Jiang H.-S., Wei T.-D., Zhang K.-Y., Wang X.-W., Zhao X.-F., Wang J.-X.: Interaction of the small GTPase Cdc42 with arginine kinase restricts white spot syndrome virus in shrimp. J Virol 2017, 91, e01916-16.

21. Xu Y., Qi Y., Luo J., Yang J., Xie Q., Deng C., Su N., Wei W., Shi D., Xu F., Li X., Xu P.: Hepatitis B virus X protein stimulates proliferation, wound closure and inhibits apoptosis of HuH-7 cells via CDC42. Int J Mol Sci 2017, 18, 586–599.

22. Yang Y.J., Liu Z.S., Lu S.Y., Li C., Hu P., Li Y.S., Liu N.N., Tang F., Xu Y.M., Zhang J.H., Li Z.H., Feng X.L., Zhou Y., Ren H.L.: Molecular cloning, expression and characterization of programmed cell death 10 from sheep (Ovis aries). Gene 2015, 558, 65–74.

Journal of Veterinary Research

formerly Bulletin of the Veterinary Institute in Pulawy

Journal Information


IMPACT FACTOR J Vet Res 2017: 0.811

CiteScore 2017: 0.68

SCImago Journal Rank (SJR) 2017: 0.29
Source Normalized Impact per Paper (SNIP) 2017: 0.484

Metrics

All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 208 208 29
PDF Downloads 73 73 7