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The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain

The underlying mechanisms by which Newcastle disease virus (NDV) kills cancer cells are still unclear. Recent discoveries have shown that many viruses contain Bcl-2 homology-like domains which enabled their interaction with Bcl-2 family members, and thereby accounting for their virulence and pathoge...

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Autores principales: Molouki, Aidin, Hsu, Yi-Te, Jahanshiri, Fatemeh, Abdullah, Syahril, Rosli, Rozita, Yusoff, Khatijah
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3166938/
https://www.ncbi.nlm.nih.gov/pubmed/21810274
http://dx.doi.org/10.1186/1743-422X-8-385
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author Molouki, Aidin
Hsu, Yi-Te
Jahanshiri, Fatemeh
Abdullah, Syahril
Rosli, Rozita
Yusoff, Khatijah
author_facet Molouki, Aidin
Hsu, Yi-Te
Jahanshiri, Fatemeh
Abdullah, Syahril
Rosli, Rozita
Yusoff, Khatijah
author_sort Molouki, Aidin
collection PubMed
description The underlying mechanisms by which Newcastle disease virus (NDV) kills cancer cells are still unclear. Recent discoveries have shown that many viruses contain Bcl-2 homology-like domains which enabled their interaction with Bcl-2 family members, and thereby accounting for their virulence and pathogenicity. Alignment of the protein sequences of Malaysian strain of NDV, known as AF2240, with those from members of the human Bcl-2 family showed many similar regions; most notably we found that its matrix (AF2240-M) protein, large (AF2240-L) protein and fusion (AF2240-F) protein all contain BH3-like regions. In addition, there are BH1-like domains in these proteins, where AF2240-F and Mcl-1 share 55% identity within this region. To further investigate our hypothesis that the presence of the BH3-like domains in these proteins may convey cytotoxicity, AF2240-M and AF2240-F genes were cloned into pFLAG and pEGFP.N2 vectors and transfected into HeLa cells. The expression of these constructs promoted cell death. As shown by flow cytometry, AF2240-M protein with deleted BH3-like region showed five-fold decrease in apoptosis. Moreover, the construct containing the N-terminal of AF2240-M showed nearly the same cell death rate as to that of the full-length protein, strongly suggesting that the BH3-like domain within this protein participates in promoting cell death. Moreover, AF2240-M transfection promoted Bax redistribution to mitochondria. Therefore, to determine whether there is any direct interaction between NDV viral proteins with some members of the Bcl-2 family, various constructs were co-transfected into HeLa cells. Co-immunoprecipitation trials showed that the AF2240-M indeed directly interacted with Bax protein via its BH3-domain, as the mutant proteins failed to interact with Bax. AF2240-F failed to interact with any of the tested proteins, although Bcl-XL slowed down the rate of cell death caused by this construct by nearly five-fold. In a parallel experiment, the level of expression of endogenous Bax and Bcl-2 after infection of HeLa cells with NDV was assessed by qRT-PCR, but no statistically significant change was observed. Consequently, the Bax/Bcl-2 ratio at the mRNA level did not alter. Overall, our study has shed additional light into the mechanisms by which NDV induces apoptosis.
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spelling pubmed-31669382011-09-06 The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain Molouki, Aidin Hsu, Yi-Te Jahanshiri, Fatemeh Abdullah, Syahril Rosli, Rozita Yusoff, Khatijah Virol J Research The underlying mechanisms by which Newcastle disease virus (NDV) kills cancer cells are still unclear. Recent discoveries have shown that many viruses contain Bcl-2 homology-like domains which enabled their interaction with Bcl-2 family members, and thereby accounting for their virulence and pathogenicity. Alignment of the protein sequences of Malaysian strain of NDV, known as AF2240, with those from members of the human Bcl-2 family showed many similar regions; most notably we found that its matrix (AF2240-M) protein, large (AF2240-L) protein and fusion (AF2240-F) protein all contain BH3-like regions. In addition, there are BH1-like domains in these proteins, where AF2240-F and Mcl-1 share 55% identity within this region. To further investigate our hypothesis that the presence of the BH3-like domains in these proteins may convey cytotoxicity, AF2240-M and AF2240-F genes were cloned into pFLAG and pEGFP.N2 vectors and transfected into HeLa cells. The expression of these constructs promoted cell death. As shown by flow cytometry, AF2240-M protein with deleted BH3-like region showed five-fold decrease in apoptosis. Moreover, the construct containing the N-terminal of AF2240-M showed nearly the same cell death rate as to that of the full-length protein, strongly suggesting that the BH3-like domain within this protein participates in promoting cell death. Moreover, AF2240-M transfection promoted Bax redistribution to mitochondria. Therefore, to determine whether there is any direct interaction between NDV viral proteins with some members of the Bcl-2 family, various constructs were co-transfected into HeLa cells. Co-immunoprecipitation trials showed that the AF2240-M indeed directly interacted with Bax protein via its BH3-domain, as the mutant proteins failed to interact with Bax. AF2240-F failed to interact with any of the tested proteins, although Bcl-XL slowed down the rate of cell death caused by this construct by nearly five-fold. In a parallel experiment, the level of expression of endogenous Bax and Bcl-2 after infection of HeLa cells with NDV was assessed by qRT-PCR, but no statistically significant change was observed. Consequently, the Bax/Bcl-2 ratio at the mRNA level did not alter. Overall, our study has shed additional light into the mechanisms by which NDV induces apoptosis. BioMed Central 2011-08-03 /pmc/articles/PMC3166938/ /pubmed/21810274 http://dx.doi.org/10.1186/1743-422X-8-385 Text en Copyright ©2011 Molouki et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Molouki, Aidin
Hsu, Yi-Te
Jahanshiri, Fatemeh
Abdullah, Syahril
Rosli, Rozita
Yusoff, Khatijah
The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain
title The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain
title_full The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain
title_fullStr The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain
title_full_unstemmed The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain
title_short The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain
title_sort matrix (m) protein of newcastle disease virus binds to human bax through its bh3 domain
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3166938/
https://www.ncbi.nlm.nih.gov/pubmed/21810274
http://dx.doi.org/10.1186/1743-422X-8-385
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