Isolation and characterisation of peroxiredoxin 6 promoter from sheep (Ovis aries)

Liu, Nan-Nan; Liu, Zeng-Shan; Lu, Shi-Ying; Hu, Pan; Zhang, Ying; Fu, Bao-Quan; Li, Yan-Song; Zhou, Yu; Zhang, Yu; Ren, Hong-Lin

doi:10.1515/jvetres-2016-0048

Abstract

Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein and a unique 1-Cys Prdx of the peroxiredoxin family. The expression and regulation of Prdx6 are implicated in numerous physiological and pathological processes.

Material and Methods: Eight stepwise truncated DNA fragments obtained from the 5′-flank region of the Prdx6 gene were prepared and subcloned into the pSEAP2-Enhancer vectors. To investigate the transcriptional activity of the truncated DNA fragments, the recombinant plasmids were transfected into the COS-1 cells and the transcriptional activity was measured via assaying the expression of the reporter gene of the secreted alkaline phosphatase.

Results: A 3.4 kb 5′-upstream flank region of the Prdx6 gene was cloned and sequenced. The region from −108 nt to −36 nt of the 5′-flanking region of the Prdx6 gene contained basal transcriptional activity.

Conclusion: This result provides the basis for further studies on the gene regulation of the Prdx6-mediated biological processes and on screening for the transacting factors that interact with cis-acting elements of the Prdx6 gene promoter.

References

1. Bast A., Erttmann S.F., Walther R., Steinmetz I.: Influence of iNOS and COX on peroxiredoxin gene expression in primary macrophages. Free Radic Biol Med 2010, 49, 1881–1891.10.1016/j.freeradbiomed.2010.09.015
Search in Google Scholar Back to article
2. Chhunchha B., Fatma N., Bhargavan B., Kubo E., Kumar A., Singh D.P.: Specificity protein, Sp1-mediated increased expression of Prdx6 as a curcumin-induced antioxidant defense in lens epithelial cells against oxidative stress. Cell Death Dis 2011, 2, e234.10.1038/cddis.2011.121
Search in Google Scholar Back to article
3. Chowdhury I., Mo Y., Gao L., Kazi A., Fisher A.B., Feinstein S.I.: Oxidant stress stimulates expression of the human peroxiredoxin 6 gene by a transcriptional mechanism involving an antioxidant response element. Free Radic Biol Med 2009, 46, 146–153.10.1016/j.freeradbiomed.2008.09.027
Search in Google Scholar Back to article
4. David E., Tanguy A., Moraga D.: Peroxiredoxin 6 gene: a new physiological and genetic indicator of multiple environmental stress response in Pacific oyster Crassostrea gigas. Aquat Toxicol 2007, 84, 389–398.10.1016/j.aquatox.2007.06.017
Search in Google Scholar Back to article
5. De Zoysa M., Ryu J.H., Chung H.C., Kim C.H., Nikapitiya C., Oh C., Kim H., Saranya Revathy K., Whang I., Lee J.: Molecular characterization, immune responses and DNA protection activity of rock bream (Oplegnathus fasciatus), peroxiredoxin 6 (Prx6). Fish Shellfish Immunol 2012, 33, 28–35.10.1016/j.fsi.2012.03.029
Search in Google Scholar Back to article
6. Fatemi M., Pao M.M., Jeong S., Gal-Yam E.N., Egger G., Weisenberger D.J., Jones P.A.: Footprinting of mammalian promoters: use of a CpG DNA methyltransferase revealing nucleosome positions at a single molecule level. Nucleic Acids Res 2005, 33, e176.10.1093/nar/gni180
Search in Google Scholar Back to article
7. Liu N.N., Liu Z.S., Lu S.Y., Hu P., Li Y.S., Feng X.L., Zhang S.Y., Wang N., Meng Q.F., Yang Y.J., Tang F., Xu Y.M., Zhang W.H., Guo X., Chen X.F., Zhou Y., Ren H.L.: Full-length cDNA cloning, molecular characterization and differential expression analysis of peroxiredoxin 6 from Ovis aries. Vet Immunol Immunopathol 2015, 164, 208–219.10.1016/j.vetimm.2015.01.006
Search in Google Scholar Back to article
8. Manevich Y., Fisher A.B.: Peroxiredoxin 6, a 1-Cys peroxiredoxin, functions in antioxidant defense and lung phospholipid metabolism. Free Radic Biol Med 2005, 38, 1422–1432.10.1016/j.freeradbiomed.2005.02.011
Search in Google Scholar Back to article
9. Nikapitiya C., De Zoysa M., Whang I., Kim C.G., Lee Y.H., Kim S.J., Lee J.: Molecular cloning, characterization and expression analysis of peroxiredoxin 6 from disk abalone Haliotis discus discus and the antioxidant activity of its recombinant protein. Fish Shellfish Immunol 2009, 27, 239–249.10.1016/j.fsi.2009.05.002
Search in Google Scholar Back to article
10. Pak J.H., Kim T.I., Joon Kim M., Yong Kim J., Choi H.J., Kim S.A., Tchah H.: Reduced expression of 1-cys peroxiredoxin in oxidative stress-induced cataracts. Exp Eye Res 2006, 82, 899–906.10.1016/j.exer.2005.10.017
Search in Google Scholar Back to article
11. Ryu H., Lee J., Zaman K., Kubilis J., Ferrante R.J., Ross B.D., Neve R., Ratan R.R.: Sp1 and Sp3 are oxidative stress-inducible, antideath transcription factors in cortical neurons. J Neurosci 2003, 23, 3597–606.10.1523/JNEUROSCI.23-09-03597.2003
Search in Google Scholar Back to article
12. Wang Q., Chen K., Yao Q., Zhao Y., Li Y., Shen H., Mu R.: Identification and characterization of a novel 1-Cys peroxiredoxin from silkworm, Bombyx mori. Comp Biochem Physiol B Biochem Mol Biol 2008, 149, 176–182.10.1016/j.cbpb.2007.09.005
Search in Google Scholar Back to article
13. Wang Y., Phelan S.A., Manevich Y., Feinstein S.I., Fisher A.B.: Transgenic mice overexpressing peroxiredoxin 6 show increased resistance to lung injury in hyperoxia. Am J Respir Cell Mol Biol 2006, 34, 481–486.10.1165/rcmb.2005-0333OC
Search in Google Scholar Back to article
14. Wu X., Ji P., Zhang L., Bu G., Gu H., Wang X., Xiong Y., Zuo B.: The expression of porcine Prdx6 gene is up-regulated by C/EBPbeta and CREB. PLoS One 2015, 10, e0144851.10.1371/journal.pone.0144851
Search in Google Scholar Back to article
15. Yun H.M., Park K.R., Lee H.P., Lee D.H., Jo M., Shin D.H., Yoon D.Y., Han S.B., Hong J.T.: PRDX6 promotes lung tumor progression via its GPx and iPLA2 activities. Free Radic Biol Med 2014, 69, 367–376.10.1016/j.freeradbiomed.2014.02.001
Search in Google Scholar Back to article
16. Zheng W.J., Hu Y.H., Zhang M., Sun L.: Analysis of the expression and antioxidative property of a peroxiredoxin 6 from Scophthalmus maximus. Fish Shellfish Immunol 2010, 29, 305–311.10.1016/j.fsi.2010.04.008
Search in Google Scholar Back to article

Isolation and characterisation of peroxiredoxin 6 promoter from sheep (Ovis aries)

Abstract

Paradigm

My account