Identification of Binding Proteins That Interact with BVDV E2 Protein in Bovine Trophoblast Cell
Bovine viral diarrhea virus (BVDV) can cause lifelong
persistent infection. One reason for the phenomena is attributed to
BVDV infection to placenta tissue. However the mechanisms that
BVDV invades into placenta tissue remain unclear. To clarify the
molecular mechanisms, we investigated the possible means that
BVDV entered into bovine trophoblast cells (TPC). Yeast two-hybrid
system was used to identify proteins extracted from TPC, which
interact with BVDV envelope glycoprotein E2. A PGbkt7-E2 yeast
expression vector and TPC cDNA library were constructed. Through
two rounds of screening, three positive clones were identified.
Sequencing analysis indicated that all the three positive clones
encoded the same protein clathrin. Physical interaction between
clathrin and BVDV E2 protein was further confirmed by
coimmunoprecipitation experiments. This result suggested that the
clathrin might play a critical role in the process of BVDV entry into
placenta tissue and might be a novel antiviral target for preventing
BVDV infection.
[1] Xiang, X., Duan, G., Li, Z. The Advance of Molecular Biology Research
of BVDV. Acta Ecologiae Animalis Domastici, 2004, 25(4): 202-204.
[2] Vidricaire, G., Tardif, M. R., Tremblay, M. J. The low viral production in
trophoblastic cells is due to a high endocytif internalization of the human
immunodeficiency virus type 1 and can be overcome by the
pro-inflammatory cytokines tumor necrosis factor-╬▒ and interleukin-1. J
biol chemistry, 2003, 278 (18): 15832-15841.
[3] Shearer, W. T., Reuben, J., Lee, B. N., Popek, E. J., Lewis, D. E.,
Hammill, H. H., Hanson, I. C., Kline, M. W., Langston, C. Role of
placental cytokines and inflammation in vertical transmission of HIV
infection. Acta Paediatr Supp, 1997, 421:33-8.
[4] Moussa, M., Mognetti, B., Dubanchet, S., Menu, E., Roques, P., Gras, G.,
Dormont, D., Barre-Sinoussi, F.,Chaouat, G. Vertical transmission of
HIV: parameters which might affect infection of placental trophoblasts by
HIV-1: a review. Biomed group on the study of in utero transmission of
HIV-1. Am J Reprod Immunol, 1999, 41(5):312-9.
[5] Parry, S., Holder, J., Strauss, J. F., Mechanisms of trophoblast-virus
interaction. J Reprod Immunol, 1997, 37(1):25-34.
[6] Gan, X., Hou, R., Zhang, M., Zhang, H. Role of Trophocytes in
Mammalian Embryo Implantation. J Sichuan Zoology, 2005, 24(4):
660-664.
[7] Ren, Y., Chen, C., Qiao, J., Wang, Y., Guo, Q., Zhang, H., Wang, P.
Culture of trophoblast giant cells from bovine placentomes. Chin J Cell
Biol, 2009, 29(7): 541-549.
[8] Wang, Z., Ding, M. Research advances on Bovine viral diarrhea virus.
Virological Sinica, 1997, 12(2): 112-118.
[9] Yu, M., Gould, A. R., Morrissy, C. J. High level expression of the
envelope glycoprotein(gp53) of bovine viral diarrhea virus (Singer) and
its potential use as diagnostic reagent. Virus Res, 1994, 34: 178-186.
[10] Donis, R. O., Corapi, W. V., Dubovi, E. J. Bovine viral diarrhea virus
proteins and their antigenic analysis. Arch Virol, 1991, Suppl 3:29-40.
[11] Jiang, X., Yang, J., Ren, K. The yeast two-hybrid system. Chin J Cell
Biol, 2000, 22(4): 176-181.
[12] Zahoor, M. A., Yamane, D., Mohamed, Y. M., Suda, Y., Kobayashi, K.,
Kato, K., Tohy,a Y., Akashi, H. Bovine viral diarrhea virus non-structural
protein 5A interacts with NIK- and IKKbeta-binding protein. J Gen Virol,
2010, 91(8):1939-48.
[13] Grummer, B., Beer, M., Liebler-Tenorio, E., Greiser-Wilke, I.
Localization of viral proteins in cells infected with bovine viral diarrhoea
virus. J Gen Virol, 2001, 82(11):2597-605.
[14] Chimeno Zoth, S., Leunda, M. R., Ode├│n, A., Taboga, O. Recombinant
E2 glycoprotein of bovine viral diarrhea virus induces a solid humoral
neutralizing immune response but fails to confer total protection in cattle.
J Virol Methods, 2006, 138(1-2):99-108.
[15] Chimeno Zoth, S., Taboga, O. Multiple recombinant ELISA for the
detection of bovine viral diarrhoea virus antibodies in cattle sera. J Virol
Methods, 2006,138(1-2):99-108.
[16] Botton, S. A., da-Silva, A. M., Brum, M. C., Weiblen, R., Flores, E. F.
Antigenic characterization of Brazilian bovine viral diarrhea virus
isolates by monoclonal antibodies and cross-neutralization. Braz J Med
Biol Res, 1998, 31(11):1429-38.
[17] Li, M. F., Shi, Y. L. The mechanism of clathrin-mediatedsynaptic vesicle
endocytosis. Chin J Neurosci, 2002, 18(3):643-647.
[18] Lecot, S., Belouzard, S., Dubuisson, J., Rouillé, Y. Bovine Viral Diarrhea
Virus Entry Is Dependent on Clathrin-Mediated Endocytosis. J Virol,
2005, 79(16):10826-9.
[19] Chu, J. J. H., and M. L. Ng. Infectious entry of West Nile virus occurs
through a clathrin-mediated endocytic pathway. J. Virol, 2004,
78:10543-10555.
[20] Grummer, B., Grotha, S. and Greiser-Wilke, I. Bovine viral diarrhea virus
is internalized by clathrin-dependent receptor-mediated endocytosis. J.
Vet. Med. B Infect. Dis. Vet. Public Health, 2004, 51:427-432.
[21] Gruenberg, J. Viruses and endosome membrane dynamics. Curr Opin
Cell Biol, 2009, 21(4):582-8.
[22] Liang, M., Morizono, K., Pariente, N., Kamata, M., Lee, B., Chen, I. S.
Targeted transduction via CD4 by a lentiviral vector uses a
clathrin-mediated entry pathway. J Virol, 2009, 83(24):13026-31.
[23] Sun, X., Yau, V. K., Briggs, B. J., Whittaker, G. R. Role of
clathrin-mediated endocytosis during vesicular stomatitis virus entry into
host cells. Virology, 2005, 338(1):53-60.
[24] Jin, M., Park, J., Lee, S., Park, B., Shin, J., Song, K. J., Ahn, T. I., Hwang,
S. Y., Ahn, B. Y., Ahn, K. Hantaan virus enters cells by
clathrin-dependent receptor-mediated endocytosis. Virology, 2002,
294(1):60-9.
[25] Querbes, W., Benmerah, A., Tosoni, D., Di Fiore, P. P., Atwood, W. J. A
JC virus-induced signal is required for infection of glial cells by a
clathrin- and eps15-dependent pathway. J Virol, 2004,78(1):250-6.
[26] Suikkanen, S., Sääjärvi, K., Hirsimäki, J., Välilehto, O., Reunanen,
H.,Vihinen-Ranta, M., Vuento, M. Role of recycling endosomes and
lysosomes in dynein-dependent entry of canine parvovirus. J Virol, 2002,
76(9):4401-11.
[27] Ruiz-Sáenz, J., Goez, Y., Tabares, W., López-Herrera, A. Cellular
receptors for foot and mouth disease virus. Intervirology, 2009,
52(4):201-12.
[28] Schmid, S. L. Clathrin coated vesicle formation and protein sorting an
integrated process. Annu Rev Biochem, 1997, 66:511-548.
[29] Pelkmans, L., Helenius, A. Endocytosis via caveolae. Traffic, 2002,
3(5):311
[30] Brodsky, F. M. Clathrin light chains: array of protein M otifs that regulate
coated-vesicle dynam ics. Trends Biology Sci, 1991, 16:208.
[31] Ihrke, G., Hubbard, A. L. Control of vesicle traffic in hepatocyte. Prog
Liver Bis, 1995, 13:63.
[32] Sle pnev, V. I., De Camilli, P. Accessory factors in clathrin dependent
synaptic vesicle endocytosis . Nat Rev Neurosci, 2000, 1:161-172.
[1] Xiang, X., Duan, G., Li, Z. The Advance of Molecular Biology Research
of BVDV. Acta Ecologiae Animalis Domastici, 2004, 25(4): 202-204.
[2] Vidricaire, G., Tardif, M. R., Tremblay, M. J. The low viral production in
trophoblastic cells is due to a high endocytif internalization of the human
immunodeficiency virus type 1 and can be overcome by the
pro-inflammatory cytokines tumor necrosis factor-╬▒ and interleukin-1. J
biol chemistry, 2003, 278 (18): 15832-15841.
[3] Shearer, W. T., Reuben, J., Lee, B. N., Popek, E. J., Lewis, D. E.,
Hammill, H. H., Hanson, I. C., Kline, M. W., Langston, C. Role of
placental cytokines and inflammation in vertical transmission of HIV
infection. Acta Paediatr Supp, 1997, 421:33-8.
[4] Moussa, M., Mognetti, B., Dubanchet, S., Menu, E., Roques, P., Gras, G.,
Dormont, D., Barre-Sinoussi, F.,Chaouat, G. Vertical transmission of
HIV: parameters which might affect infection of placental trophoblasts by
HIV-1: a review. Biomed group on the study of in utero transmission of
HIV-1. Am J Reprod Immunol, 1999, 41(5):312-9.
[5] Parry, S., Holder, J., Strauss, J. F., Mechanisms of trophoblast-virus
interaction. J Reprod Immunol, 1997, 37(1):25-34.
[6] Gan, X., Hou, R., Zhang, M., Zhang, H. Role of Trophocytes in
Mammalian Embryo Implantation. J Sichuan Zoology, 2005, 24(4):
660-664.
[7] Ren, Y., Chen, C., Qiao, J., Wang, Y., Guo, Q., Zhang, H., Wang, P.
Culture of trophoblast giant cells from bovine placentomes. Chin J Cell
Biol, 2009, 29(7): 541-549.
[8] Wang, Z., Ding, M. Research advances on Bovine viral diarrhea virus.
Virological Sinica, 1997, 12(2): 112-118.
[9] Yu, M., Gould, A. R., Morrissy, C. J. High level expression of the
envelope glycoprotein(gp53) of bovine viral diarrhea virus (Singer) and
its potential use as diagnostic reagent. Virus Res, 1994, 34: 178-186.
[10] Donis, R. O., Corapi, W. V., Dubovi, E. J. Bovine viral diarrhea virus
proteins and their antigenic analysis. Arch Virol, 1991, Suppl 3:29-40.
[11] Jiang, X., Yang, J., Ren, K. The yeast two-hybrid system. Chin J Cell
Biol, 2000, 22(4): 176-181.
[12] Zahoor, M. A., Yamane, D., Mohamed, Y. M., Suda, Y., Kobayashi, K.,
Kato, K., Tohy,a Y., Akashi, H. Bovine viral diarrhea virus non-structural
protein 5A interacts with NIK- and IKKbeta-binding protein. J Gen Virol,
2010, 91(8):1939-48.
[13] Grummer, B., Beer, M., Liebler-Tenorio, E., Greiser-Wilke, I.
Localization of viral proteins in cells infected with bovine viral diarrhoea
virus. J Gen Virol, 2001, 82(11):2597-605.
[14] Chimeno Zoth, S., Leunda, M. R., Ode├│n, A., Taboga, O. Recombinant
E2 glycoprotein of bovine viral diarrhea virus induces a solid humoral
neutralizing immune response but fails to confer total protection in cattle.
J Virol Methods, 2006, 138(1-2):99-108.
[15] Chimeno Zoth, S., Taboga, O. Multiple recombinant ELISA for the
detection of bovine viral diarrhoea virus antibodies in cattle sera. J Virol
Methods, 2006,138(1-2):99-108.
[16] Botton, S. A., da-Silva, A. M., Brum, M. C., Weiblen, R., Flores, E. F.
Antigenic characterization of Brazilian bovine viral diarrhea virus
isolates by monoclonal antibodies and cross-neutralization. Braz J Med
Biol Res, 1998, 31(11):1429-38.
[17] Li, M. F., Shi, Y. L. The mechanism of clathrin-mediatedsynaptic vesicle
endocytosis. Chin J Neurosci, 2002, 18(3):643-647.
[18] Lecot, S., Belouzard, S., Dubuisson, J., Rouillé, Y. Bovine Viral Diarrhea
Virus Entry Is Dependent on Clathrin-Mediated Endocytosis. J Virol,
2005, 79(16):10826-9.
[19] Chu, J. J. H., and M. L. Ng. Infectious entry of West Nile virus occurs
through a clathrin-mediated endocytic pathway. J. Virol, 2004,
78:10543-10555.
[20] Grummer, B., Grotha, S. and Greiser-Wilke, I. Bovine viral diarrhea virus
is internalized by clathrin-dependent receptor-mediated endocytosis. J.
Vet. Med. B Infect. Dis. Vet. Public Health, 2004, 51:427-432.
[21] Gruenberg, J. Viruses and endosome membrane dynamics. Curr Opin
Cell Biol, 2009, 21(4):582-8.
[22] Liang, M., Morizono, K., Pariente, N., Kamata, M., Lee, B., Chen, I. S.
Targeted transduction via CD4 by a lentiviral vector uses a
clathrin-mediated entry pathway. J Virol, 2009, 83(24):13026-31.
[23] Sun, X., Yau, V. K., Briggs, B. J., Whittaker, G. R. Role of
clathrin-mediated endocytosis during vesicular stomatitis virus entry into
host cells. Virology, 2005, 338(1):53-60.
[24] Jin, M., Park, J., Lee, S., Park, B., Shin, J., Song, K. J., Ahn, T. I., Hwang,
S. Y., Ahn, B. Y., Ahn, K. Hantaan virus enters cells by
clathrin-dependent receptor-mediated endocytosis. Virology, 2002,
294(1):60-9.
[25] Querbes, W., Benmerah, A., Tosoni, D., Di Fiore, P. P., Atwood, W. J. A
JC virus-induced signal is required for infection of glial cells by a
clathrin- and eps15-dependent pathway. J Virol, 2004,78(1):250-6.
[26] Suikkanen, S., Sääjärvi, K., Hirsimäki, J., Välilehto, O., Reunanen,
H.,Vihinen-Ranta, M., Vuento, M. Role of recycling endosomes and
lysosomes in dynein-dependent entry of canine parvovirus. J Virol, 2002,
76(9):4401-11.
[27] Ruiz-Sáenz, J., Goez, Y., Tabares, W., López-Herrera, A. Cellular
receptors for foot and mouth disease virus. Intervirology, 2009,
52(4):201-12.
[28] Schmid, S. L. Clathrin coated vesicle formation and protein sorting an
integrated process. Annu Rev Biochem, 1997, 66:511-548.
[29] Pelkmans, L., Helenius, A. Endocytosis via caveolae. Traffic, 2002,
3(5):311
[30] Brodsky, F. M. Clathrin light chains: array of protein M otifs that regulate
coated-vesicle dynam ics. Trends Biology Sci, 1991, 16:208.
[31] Ihrke, G., Hubbard, A. L. Control of vesicle traffic in hepatocyte. Prog
Liver Bis, 1995, 13:63.
[32] Sle pnev, V. I., De Camilli, P. Accessory factors in clathrin dependent
synaptic vesicle endocytosis . Nat Rev Neurosci, 2000, 1:161-172.
@article{"International Journal of Biological, Life and Agricultural Sciences:56182", author = "Yan Ren and Fei Guo and Jun Qiao and Shengwei Hu and Hui Zhang and Yuanzhi Wang and Pengyan Wang and Jinliang Sheng and Xinli Gu and Xiaojun Liu and Chuangfu Chen", title = "Identification of Binding Proteins That Interact with BVDV E2 Protein in Bovine Trophoblast Cell", abstract = "Bovine viral diarrhea virus (BVDV) can cause lifelong
persistent infection. One reason for the phenomena is attributed to
BVDV infection to placenta tissue. However the mechanisms that
BVDV invades into placenta tissue remain unclear. To clarify the
molecular mechanisms, we investigated the possible means that
BVDV entered into bovine trophoblast cells (TPC). Yeast two-hybrid
system was used to identify proteins extracted from TPC, which
interact with BVDV envelope glycoprotein E2. A PGbkt7-E2 yeast
expression vector and TPC cDNA library were constructed. Through
two rounds of screening, three positive clones were identified.
Sequencing analysis indicated that all the three positive clones
encoded the same protein clathrin. Physical interaction between
clathrin and BVDV E2 protein was further confirmed by
coimmunoprecipitation experiments. This result suggested that the
clathrin might play a critical role in the process of BVDV entry into
placenta tissue and might be a novel antiviral target for preventing
BVDV infection.", keywords = "Bovine viral diarrhea virus, clathrin, glycoprotein E2, yeast two-hybrid system.", volume = "7", number = "3", pages = "202-4", }
