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Research Article

volume 53 issue 6 (june 2019) : 724-730,   Doi: 10.18805/ijar.B-883

Effect of TRIM28 on proliferation, apoptosis and histone H3K9 trimethylation in bovine fibroblasts

P
Panpan Ma
X
Xiaxia Man
S
Shubao Yang
X
Xiaoqing Dong
L
Liyan Su
W
Weimin Luan
X
Xin Ma
1College of Animal Science and Technology, Jilin Agricultural University, Changchun 130 118, China.
Cite article:- Ma Panpan, Man Xiaxia, Yang Shubao, Dong Xiaoqing, Su Liyan, Luan Weimin, Ma Xin (2019). Effect of TRIM28 on proliferation, apoptosis and histone H3K9 trimethylation in bovine fibroblasts. Indian Journal of Animal Research. 53(6): 724-730. doi: 10.18805/ijar.B-883.

ABSTRACT

TRIM28 is a co-repressor, which interacts with HP1 proteins and chromatin repressive complexes leading to gene expression silencing. In this study, after we used Lipofectamine3000-mediating siRNA and restructured p EGFP-IRES2-TRIM28 to transfect bovine fibroblasts, the expression levels of TRIM28, HP1BP3, DNMT, SETDB1, TP53, BAX, Bcl and SOD were detected by real-time polymerase chain reaction (PCR). In addition, the influence of p EGFP-IRES2-TRIM28 on histone H3K9me3 was also observed. In the siRNA transfected group, TRIM28 was down-regulated (P<0.01), which inhibited cell proliferation and reduced the level of H3K9me3 expression. In the p EGFP-IRES2-TRIM28 transfected group, TRIM28 was over-expressed (P<0.01), and the pro-apoptosis gene BAX was significantly decreased (P<0.01), but there was no significant change in other genes. Our results demonstrate that TRIM28 plays an important role in bovine fibroblasts and might be a valuable biomolecule for proliferation, apoptosis, and histone H3K9 trimethylation.

REFERENCES

  1. Babu, P. P., Suzuki, G., Ono, Y. and Yoshida, Y. (2004). Attenuation of ischemia and/or reperfusion injury during myocardial infarction using mild hypothermia in rats: an immunohistochemical study of bcl-2, BAX, bak and tunel. Pathology International, 54:896–903. 
  2. Elbashir, S. M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K. and Tuschl, T. (2001). Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature, 411:494–498.
  3. Friedman, J.R., Fredericks, W.J., Jensen, D.E., Speicher, D. W., Huang, X. P. and Neilson, E.G. (1996). Kap-1, a novel corepressor for the highly conserved KRAB repression domain. Genes and Development, 10(16):2067-78.
  4. Fire, A., Xu, S., Montgomery, M.K., Kostas, S.A., Driver, S.E. and Mello, C.C. (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391:806–811.
  5. Hamm, J., Tessanne, K., Murphy, C.N. and Prather, R.S. (2014). Transcriptional regulators trim28, setdb1, and tp53 are aberrantly expressed in porcine embryos produced by in vitro fertilization in comparison to in vivo- and somatic-cell nuclear transfer-    derived embryos. Molecular Reproduction & Development, 81:552.
  6. Hamadou, W. S., Besbes, S., Bourdon, V., Youssef, Y. B., Laatiri, M.A., Noguchi, T., et al. (2016). Mutational analysis of tp53 gene in Tunisian familial hematological malignancies and sporadic acute leukemia cases. Familial Cancer, 16:153-157.
  7. Hatakeyama S. (2011). TRIM proteins and cancer. Nature Reviews Cancer, 11:792–804. 
  8. Hublitz, P., Albert, M. and Peters, A.H. (2009). Mechanisms of transcriptional repression by histone lysine methylation. International Journal of Developmental Biology, 53(2-3):335.
  9. Iyengar, S. and Farnham, P. J. (2011). Kap1 protein: an enigmatic master regulator of the genome. Journal of Biological Chemistry, 286(30):26267-76.
  10. Ma, X., Zhai, Z. C., Zhang, M. L., Song, B. H., Zhu, Y. R., Yang, S. B., et al (2016). Molecular cloning and expression vector construction of bovine trim 28. Genetics & Molecular Research, 15.
  11. Nielsen, A.L., Ortiz, J.A., You, J., Oulad-Abdelghani, M., Khechumian, R. and Gansmuller, A., et al. (1999). Interaction with members of the heterochromatin protein 1 (hp1) family and histone deacetylation are differentially involved in transcriptional silencing by members of the tif1 family. EMBO Journal, 18: 6385-95.
  12. Pathak, K. and Gogoi, B. (2016). RNA interference (RNAi): Application in crop improvement: A review. Agricultural Reviews. 37:245-249.
  13. Prabhavathy, H. and Palanivel, K.M. (2015). Ability of Indian street rabies virus isolates to induce apoptosis by in-vitro. Indian Journal of Animal Research, 49: 323-46.
  14. Quenneville, S., Verde, G., Corsinotti, A., Kapopoulou, A., Jakobsson, J., Offner, S., Baglivo I, Pedone PV, Grimaldi G, Riccio A and Trono D. (2011). In embryonic stem cells, ZFP57/KAP1 recognize a methylated hexanucleotide to affect chromatin and DNA methylation of imprinting control regions. Molecular Cell, 44:361-372.
  15. Rosenfeld, J.A., Wang, Z., Schones, D0.E., Zhao, K., Desalle, R. and Zhang, M.Q. (2009). Determination of enriched histone modifications in non-genic portions of the human genome. BMC Genomics, 10:143.
  16. Schultz, D.C., Ayyanathan, K., Negorev, D., Maul, G.G. and Rauscher, F.J. (2002). SETDB1: a novel KAP-1-associated histone H3, lysine 9-specific methyltransferase that contributes to HP1-mediated silencing of euchromatic genes by KRAB zinc-finger proteins. Genes & Development, 16:919-32.
  17. Schultz, D.C., Friedman, J.R. and Rauscher, F.J. (2001). Targeting histone deacetylase complexes via KRAB-zinc finger proteins: the PHD and bromodomains of KAP-1form a cooperative unit that recruits a novel isoform of the Mi-2alpha subunit of NuRD. Genes & Development, 15:428-43.
  18. Singh, S. and Ganguli, I. (2013). RNA interference technology: implications and applications - a review. Agricultural Reviews. 34: 62-70
  19. Su, J., Wang, Y., Xing, X., Zhang, L., Sun, H. and Zhang, Y. (2015). Melatonin significantly improves the developmental competence of bovine somatic cell nuclear transfer embryos. Journal of Pineal Research, 59:455.
  20. Vakoc, C.R., Mandat, S.A., Olenchock, B.A. and Blobel, G.A. (2005). Histone h3 lysine 9 methylation and hp1gamma are associated with transcription elongation through mammalian chromatin. Molecular Cell, 19:381.
  21. Wang, Y., Jiang, J., Li, Q., Ma, H., Xu, Z. And Gao, Y. (2016). Kap1 is overexpressed in hepatocellular carcinoma and its clinical significance. International Journal of Clinical Oncology, 21:1-7.
  22. Zhang, S., Tang, B., Fan, C., Shi, L., Zhang, X., Sun, L. and Li, Z.Y. (2015). Effect of DNMT inhibitor on bovine parthenogenetic embryo development. Biochemical and Biophysical Research Communications, 466(3):505-11.
  23. Zuo, X., Sheng, J., Lau, H. T., Mcdonald, C.M., Andrade, M. and Cullen, D. E. (2012). Zinc finger protein zfp57 requires its co-factor to recruit DNA methyltransferases and maintains DNA methylation imprint in embryonic stem cells via its transcriptional repression domain. Journal of Biological Chemistry, 287(3):2107-18. 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 6 (june 2019) : 724-730,   Doi: 10.18805/ijar.B-883

    Effect of TRIM28 on proliferation, apoptosis and histone H3K9 trimethylation in bovine fibroblasts

    P
    Panpan Ma
    X
    Xiaxia Man
    S
    Shubao Yang
    X
    Xiaoqing Dong
    L
    Liyan Su
    W
    Weimin Luan
    X
    Xin Ma
    1College of Animal Science and Technology, Jilin Agricultural University, Changchun 130 118, China.
    Cite article:- Ma Panpan, Man Xiaxia, Yang Shubao, Dong Xiaoqing, Su Liyan, Luan Weimin, Ma Xin (2019). Effect of TRIM28 on proliferation, apoptosis and histone H3K9 trimethylation in bovine fibroblasts. Indian Journal of Animal Research. 53(6): 724-730. doi: 10.18805/ijar.B-883.

    ABSTRACT

    TRIM28 is a co-repressor, which interacts with HP1 proteins and chromatin repressive complexes leading to gene expression silencing. In this study, after we used Lipofectamine3000-mediating siRNA and restructured p EGFP-IRES2-TRIM28 to transfect bovine fibroblasts, the expression levels of TRIM28, HP1BP3, DNMT, SETDB1, TP53, BAX, Bcl and SOD were detected by real-time polymerase chain reaction (PCR). In addition, the influence of p EGFP-IRES2-TRIM28 on histone H3K9me3 was also observed. In the siRNA transfected group, TRIM28 was down-regulated (P<0.01), which inhibited cell proliferation and reduced the level of H3K9me3 expression. In the p EGFP-IRES2-TRIM28 transfected group, TRIM28 was over-expressed (P<0.01), and the pro-apoptosis gene BAX was significantly decreased (P<0.01), but there was no significant change in other genes. Our results demonstrate that TRIM28 plays an important role in bovine fibroblasts and might be a valuable biomolecule for proliferation, apoptosis, and histone H3K9 trimethylation.

    REFERENCES

    1. Babu, P. P., Suzuki, G., Ono, Y. and Yoshida, Y. (2004). Attenuation of ischemia and/or reperfusion injury during myocardial infarction using mild hypothermia in rats: an immunohistochemical study of bcl-2, BAX, bak and tunel. Pathology International, 54:896–903. 
    2. Elbashir, S. M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K. and Tuschl, T. (2001). Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature, 411:494–498.
    3. Friedman, J.R., Fredericks, W.J., Jensen, D.E., Speicher, D. W., Huang, X. P. and Neilson, E.G. (1996). Kap-1, a novel corepressor for the highly conserved KRAB repression domain. Genes and Development, 10(16):2067-78.
    4. Fire, A., Xu, S., Montgomery, M.K., Kostas, S.A., Driver, S.E. and Mello, C.C. (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391:806–811.
    5. Hamm, J., Tessanne, K., Murphy, C.N. and Prather, R.S. (2014). Transcriptional regulators trim28, setdb1, and tp53 are aberrantly expressed in porcine embryos produced by in vitro fertilization in comparison to in vivo- and somatic-cell nuclear transfer-    derived embryos. Molecular Reproduction & Development, 81:552.
    6. Hamadou, W. S., Besbes, S., Bourdon, V., Youssef, Y. B., Laatiri, M.A., Noguchi, T., et al. (2016). Mutational analysis of tp53 gene in Tunisian familial hematological malignancies and sporadic acute leukemia cases. Familial Cancer, 16:153-157.
    7. Hatakeyama S. (2011). TRIM proteins and cancer. Nature Reviews Cancer, 11:792–804. 
    8. Hublitz, P., Albert, M. and Peters, A.H. (2009). Mechanisms of transcriptional repression by histone lysine methylation. International Journal of Developmental Biology, 53(2-3):335.
    9. Iyengar, S. and Farnham, P. J. (2011). Kap1 protein: an enigmatic master regulator of the genome. Journal of Biological Chemistry, 286(30):26267-76.
    10. Ma, X., Zhai, Z. C., Zhang, M. L., Song, B. H., Zhu, Y. R., Yang, S. B., et al (2016). Molecular cloning and expression vector construction of bovine trim 28. Genetics & Molecular Research, 15.
    11. Nielsen, A.L., Ortiz, J.A., You, J., Oulad-Abdelghani, M., Khechumian, R. and Gansmuller, A., et al. (1999). Interaction with members of the heterochromatin protein 1 (hp1) family and histone deacetylation are differentially involved in transcriptional silencing by members of the tif1 family. EMBO Journal, 18: 6385-95.
    12. Pathak, K. and Gogoi, B. (2016). RNA interference (RNAi): Application in crop improvement: A review. Agricultural Reviews. 37:245-249.
    13. Prabhavathy, H. and Palanivel, K.M. (2015). Ability of Indian street rabies virus isolates to induce apoptosis by in-vitro. Indian Journal of Animal Research, 49: 323-46.
    14. Quenneville, S., Verde, G., Corsinotti, A., Kapopoulou, A., Jakobsson, J., Offner, S., Baglivo I, Pedone PV, Grimaldi G, Riccio A and Trono D. (2011). In embryonic stem cells, ZFP57/KAP1 recognize a methylated hexanucleotide to affect chromatin and DNA methylation of imprinting control regions. Molecular Cell, 44:361-372.
    15. Rosenfeld, J.A., Wang, Z., Schones, D0.E., Zhao, K., Desalle, R. and Zhang, M.Q. (2009). Determination of enriched histone modifications in non-genic portions of the human genome. BMC Genomics, 10:143.
    16. Schultz, D.C., Ayyanathan, K., Negorev, D., Maul, G.G. and Rauscher, F.J. (2002). SETDB1: a novel KAP-1-associated histone H3, lysine 9-specific methyltransferase that contributes to HP1-mediated silencing of euchromatic genes by KRAB zinc-finger proteins. Genes & Development, 16:919-32.
    17. Schultz, D.C., Friedman, J.R. and Rauscher, F.J. (2001). Targeting histone deacetylase complexes via KRAB-zinc finger proteins: the PHD and bromodomains of KAP-1form a cooperative unit that recruits a novel isoform of the Mi-2alpha subunit of NuRD. Genes & Development, 15:428-43.
    18. Singh, S. and Ganguli, I. (2013). RNA interference technology: implications and applications - a review. Agricultural Reviews. 34: 62-70
    19. Su, J., Wang, Y., Xing, X., Zhang, L., Sun, H. and Zhang, Y. (2015). Melatonin significantly improves the developmental competence of bovine somatic cell nuclear transfer embryos. Journal of Pineal Research, 59:455.
    20. Vakoc, C.R., Mandat, S.A., Olenchock, B.A. and Blobel, G.A. (2005). Histone h3 lysine 9 methylation and hp1gamma are associated with transcription elongation through mammalian chromatin. Molecular Cell, 19:381.
    21. Wang, Y., Jiang, J., Li, Q., Ma, H., Xu, Z. And Gao, Y. (2016). Kap1 is overexpressed in hepatocellular carcinoma and its clinical significance. International Journal of Clinical Oncology, 21:1-7.
    22. Zhang, S., Tang, B., Fan, C., Shi, L., Zhang, X., Sun, L. and Li, Z.Y. (2015). Effect of DNMT inhibitor on bovine parthenogenetic embryo development. Biochemical and Biophysical Research Communications, 466(3):505-11.
    23. Zuo, X., Sheng, J., Lau, H. T., Mcdonald, C.M., Andrade, M. and Cullen, D. E. (2012). Zinc finger protein zfp57 requires its co-factor to recruit DNA methyltransferases and maintains DNA methylation imprint in embryonic stem cells via its transcriptional repression domain. Journal of Biological Chemistry, 287(3):2107-18. 
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