Bluetongue virus infection induces aberrant mitosis in mammalian cells

Andrew E. Shaw, Anke Brüning-Richardson, Ewan E. Morrison, Jacquelyn Bond, Jennifer Simpson, Natalie Ross-Smith, Oya Alpar, Peter P.C. Mertens, Paul Monaghan

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background: Bluetongue virus (BTV) is an arbovirus that is responsible for 'bluetongue', an economically important disease of livestock. Although BTV is well characterised at the protein level, less is known regarding its interaction with host cells. During studies of virus inclusion body formation we observed what appeared to be a large proportion of cells in mitosis. Although the modulation of the cell cycle is well established for many viruses, this was a novel observation for BTV. We therefore undertook a study to reveal in more depth the impact of BTV upon cell division. Methods. We used a confocal microscopy approach to investigate the localisation of BTV proteins in a cellular context with their respective position relative to cellular proteins. In addition, to quantitatively assess the frequency of aberrant mitosis induction by the viral non-structural protein (NS) 2 we utilised live cell imaging to monitor HeLa-mCherry tubulin cells transfected with a plasmid expressing NS2. Results: Our data showed that these 'aberrant mitoses' can be induced in multiple cell types and by different strains of BTV. Further study confirmed multiplication of the centrosomes, each resulting in a separate mitotic spindle during mitosis. Interestingly, the BTV NS1 protein was strongly localised to the centrosomal regions. In a separate, yet related observation, the BTV NS2 protein was co-localised with the condensed chromosomes to a region suggestive of the kinetochore. Live cell imaging revealed that expression of an EGFP-NS2 fusion protein in HeLa-mCherry tubulin cells also results in mitotic defects. Conclusions: We hypothesise that NS2 is a microtubule cargo protein that may inadvertently disrupt the interaction of microtubule tips with the kinetochores during mitosis. Furthermore, the BTV NS1 protein was distinctly localised to a region encompassing the centrosome and may therefore be, at least in part, responsible for the disruption of the centrosome as observed in BTV infected mammalian cells.

LanguageEnglish
Article number319
JournalVirology Journal
Volume10
Early online date28 Oct 2013
DOIs
Publication statusPublished - 30 Oct 2013
Externally publishedYes

Fingerprint

Bluetongue virus
Virus Diseases
Mitosis
Centrosome
Kinetochores
Tubulin
Proteins
Viral Nonstructural Proteins
Bluetongue
Observation
Viruses
Microtubule Proteins
Arboviruses
Spindle Apparatus
Inclusion Bodies
Livestock
Confocal Microscopy
Microtubules
Cell Division
Cell Cycle

Cite this

Shaw, A. E., Brüning-Richardson, A., Morrison, E. E., Bond, J., Simpson, J., Ross-Smith, N., ... Monaghan, P. (2013). Bluetongue virus infection induces aberrant mitosis in mammalian cells. Virology Journal, 10, [319]. https://doi.org/10.1186/1743-422X-10-319
Shaw, Andrew E. ; Brüning-Richardson, Anke ; Morrison, Ewan E. ; Bond, Jacquelyn ; Simpson, Jennifer ; Ross-Smith, Natalie ; Alpar, Oya ; Mertens, Peter P.C. ; Monaghan, Paul. / Bluetongue virus infection induces aberrant mitosis in mammalian cells. In: Virology Journal. 2013 ; Vol. 10.
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Shaw, AE, Brüning-Richardson, A, Morrison, EE, Bond, J, Simpson, J, Ross-Smith, N, Alpar, O, Mertens, PPC & Monaghan, P 2013, 'Bluetongue virus infection induces aberrant mitosis in mammalian cells', Virology Journal, vol. 10, 319. https://doi.org/10.1186/1743-422X-10-319

Bluetongue virus infection induces aberrant mitosis in mammalian cells. / Shaw, Andrew E.; Brüning-Richardson, Anke; Morrison, Ewan E.; Bond, Jacquelyn; Simpson, Jennifer; Ross-Smith, Natalie; Alpar, Oya; Mertens, Peter P.C.; Monaghan, Paul.

In: Virology Journal, Vol. 10, 319, 30.10.2013.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Bluetongue virus infection induces aberrant mitosis in mammalian cells

AU - Shaw, Andrew E.

AU - Brüning-Richardson, Anke

AU - Morrison, Ewan E.

AU - Bond, Jacquelyn

AU - Simpson, Jennifer

AU - Ross-Smith, Natalie

AU - Alpar, Oya

AU - Mertens, Peter P.C.

AU - Monaghan, Paul

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N2 - Background: Bluetongue virus (BTV) is an arbovirus that is responsible for 'bluetongue', an economically important disease of livestock. Although BTV is well characterised at the protein level, less is known regarding its interaction with host cells. During studies of virus inclusion body formation we observed what appeared to be a large proportion of cells in mitosis. Although the modulation of the cell cycle is well established for many viruses, this was a novel observation for BTV. We therefore undertook a study to reveal in more depth the impact of BTV upon cell division. Methods. We used a confocal microscopy approach to investigate the localisation of BTV proteins in a cellular context with their respective position relative to cellular proteins. In addition, to quantitatively assess the frequency of aberrant mitosis induction by the viral non-structural protein (NS) 2 we utilised live cell imaging to monitor HeLa-mCherry tubulin cells transfected with a plasmid expressing NS2. Results: Our data showed that these 'aberrant mitoses' can be induced in multiple cell types and by different strains of BTV. Further study confirmed multiplication of the centrosomes, each resulting in a separate mitotic spindle during mitosis. Interestingly, the BTV NS1 protein was strongly localised to the centrosomal regions. In a separate, yet related observation, the BTV NS2 protein was co-localised with the condensed chromosomes to a region suggestive of the kinetochore. Live cell imaging revealed that expression of an EGFP-NS2 fusion protein in HeLa-mCherry tubulin cells also results in mitotic defects. Conclusions: We hypothesise that NS2 is a microtubule cargo protein that may inadvertently disrupt the interaction of microtubule tips with the kinetochores during mitosis. Furthermore, the BTV NS1 protein was distinctly localised to a region encompassing the centrosome and may therefore be, at least in part, responsible for the disruption of the centrosome as observed in BTV infected mammalian cells.

AB - Background: Bluetongue virus (BTV) is an arbovirus that is responsible for 'bluetongue', an economically important disease of livestock. Although BTV is well characterised at the protein level, less is known regarding its interaction with host cells. During studies of virus inclusion body formation we observed what appeared to be a large proportion of cells in mitosis. Although the modulation of the cell cycle is well established for many viruses, this was a novel observation for BTV. We therefore undertook a study to reveal in more depth the impact of BTV upon cell division. Methods. We used a confocal microscopy approach to investigate the localisation of BTV proteins in a cellular context with their respective position relative to cellular proteins. In addition, to quantitatively assess the frequency of aberrant mitosis induction by the viral non-structural protein (NS) 2 we utilised live cell imaging to monitor HeLa-mCherry tubulin cells transfected with a plasmid expressing NS2. Results: Our data showed that these 'aberrant mitoses' can be induced in multiple cell types and by different strains of BTV. Further study confirmed multiplication of the centrosomes, each resulting in a separate mitotic spindle during mitosis. Interestingly, the BTV NS1 protein was strongly localised to the centrosomal regions. In a separate, yet related observation, the BTV NS2 protein was co-localised with the condensed chromosomes to a region suggestive of the kinetochore. Live cell imaging revealed that expression of an EGFP-NS2 fusion protein in HeLa-mCherry tubulin cells also results in mitotic defects. Conclusions: We hypothesise that NS2 is a microtubule cargo protein that may inadvertently disrupt the interaction of microtubule tips with the kinetochores during mitosis. Furthermore, the BTV NS1 protein was distinctly localised to a region encompassing the centrosome and may therefore be, at least in part, responsible for the disruption of the centrosome as observed in BTV infected mammalian cells.

KW - Bluetongue

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KW - Non-structural

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