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Characterization of aggregate/aggresome structures formed by polyhedrin of Bombyx mori nucleopolyhedrovirus.

Guo ZJ, Tao LX, Dong XY, Yu MH, Tian T, Tang XD - Sci Rep (2015)

Bottom Line: At 48 h p.i. recovered polyhedrin bound directly to Bombyx mori microtubule-associated protein 1-light chain 3 (BmLC3), an autophagosome marker, and was colocalized with BmLC3 to the isolation membrane of autophagosome, implying the involvement of polyhedrin in cellular autophagy.Inhibition of autophagy by 3-methyladenine (3-MA) dramatically resulted in decrease of polyhedrin expression and polyhedra particle production.These observations suggested that highly expressed polyhedrin forms aggregate to get involved in cellular autophagy then play an important role in polyhedra production.

View Article: PubMed Central - PubMed

Affiliation: Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang 212013, Jiangsu, P.R. China.

ABSTRACT
Virus infections often lead to formation of aggregates and aggresomes in host cells. In this study, production of aggregates and aggresomes by the highly expressed protein polyhedrin of Bombyx mori nucleopolyhedrovirus (BmNPV) at 24 h postinfection (p.i.) was detected with a fluorescent molecular dye, and verified by colocalization of polyhedrin with aggresomal markers, GFP-250 and γ-tubulin. Polyhedrin aggregates showed hallmark characteristics of aggresomes: formation was microtubule-dependent; they colocalized with heat shock cognates/proteins of the 70-kDa family (HSC/HSP70s), ubiquitinated proteins and recruited the mitochondria. Aggregated polyhedrin protein gradually gained its active conformation accompanying progress of BmNPV infection. At 48 h p.i. recovered polyhedrin bound directly to Bombyx mori microtubule-associated protein 1-light chain 3 (BmLC3), an autophagosome marker, and was colocalized with BmLC3 to the isolation membrane of autophagosome, implying the involvement of polyhedrin in cellular autophagy. Inhibition of autophagy by 3-methyladenine (3-MA) dramatically resulted in decrease of polyhedrin expression and polyhedra particle production. These observations suggested that highly expressed polyhedrin forms aggregate to get involved in cellular autophagy then play an important role in polyhedra production.

No MeSH data available.


Related in: MedlinePlus

Protein aggregation detected by ProteoStat® dye.BmN cells were either mock- or infected with BmNPV BVs at an MOI of 10 TCID50/cell. After treatment, cells were incubated with ProteoStat® dye for 30 min. Cell nuclei were stained with Hoechst 33342. Images were obtained using a Leica TCS SP5 confocal laser-scanning microscope.
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f1: Protein aggregation detected by ProteoStat® dye.BmN cells were either mock- or infected with BmNPV BVs at an MOI of 10 TCID50/cell. After treatment, cells were incubated with ProteoStat® dye for 30 min. Cell nuclei were stained with Hoechst 33342. Images were obtained using a Leica TCS SP5 confocal laser-scanning microscope.

Mentions: ProteoStat Aggresome Detection Kit has been used extensively to specifically detect denatured and/or misfolded protein aggregates and inclusion bodies in Escherichia coli, plant and mammalian cells17181920212223. During BmNPV infection, protein aggregates/aggresomes (foci) were detected by this dye to be localized largely in the cytoplasm. Some dots that represents polyhedral occlusion bodies (OBs), were also observed in the nucleus at 48 h p.i. (Fig. 1).


Characterization of aggregate/aggresome structures formed by polyhedrin of Bombyx mori nucleopolyhedrovirus.

Guo ZJ, Tao LX, Dong XY, Yu MH, Tian T, Tang XD - Sci Rep (2015)

Protein aggregation detected by ProteoStat® dye.BmN cells were either mock- or infected with BmNPV BVs at an MOI of 10 TCID50/cell. After treatment, cells were incubated with ProteoStat® dye for 30 min. Cell nuclei were stained with Hoechst 33342. Images were obtained using a Leica TCS SP5 confocal laser-scanning microscope.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4594129&req=5

f1: Protein aggregation detected by ProteoStat® dye.BmN cells were either mock- or infected with BmNPV BVs at an MOI of 10 TCID50/cell. After treatment, cells were incubated with ProteoStat® dye for 30 min. Cell nuclei were stained with Hoechst 33342. Images were obtained using a Leica TCS SP5 confocal laser-scanning microscope.
Mentions: ProteoStat Aggresome Detection Kit has been used extensively to specifically detect denatured and/or misfolded protein aggregates and inclusion bodies in Escherichia coli, plant and mammalian cells17181920212223. During BmNPV infection, protein aggregates/aggresomes (foci) were detected by this dye to be localized largely in the cytoplasm. Some dots that represents polyhedral occlusion bodies (OBs), were also observed in the nucleus at 48 h p.i. (Fig. 1).

Bottom Line: At 48 h p.i. recovered polyhedrin bound directly to Bombyx mori microtubule-associated protein 1-light chain 3 (BmLC3), an autophagosome marker, and was colocalized with BmLC3 to the isolation membrane of autophagosome, implying the involvement of polyhedrin in cellular autophagy.Inhibition of autophagy by 3-methyladenine (3-MA) dramatically resulted in decrease of polyhedrin expression and polyhedra particle production.These observations suggested that highly expressed polyhedrin forms aggregate to get involved in cellular autophagy then play an important role in polyhedra production.

View Article: PubMed Central - PubMed

Affiliation: Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang 212013, Jiangsu, P.R. China.

ABSTRACT
Virus infections often lead to formation of aggregates and aggresomes in host cells. In this study, production of aggregates and aggresomes by the highly expressed protein polyhedrin of Bombyx mori nucleopolyhedrovirus (BmNPV) at 24 h postinfection (p.i.) was detected with a fluorescent molecular dye, and verified by colocalization of polyhedrin with aggresomal markers, GFP-250 and γ-tubulin. Polyhedrin aggregates showed hallmark characteristics of aggresomes: formation was microtubule-dependent; they colocalized with heat shock cognates/proteins of the 70-kDa family (HSC/HSP70s), ubiquitinated proteins and recruited the mitochondria. Aggregated polyhedrin protein gradually gained its active conformation accompanying progress of BmNPV infection. At 48 h p.i. recovered polyhedrin bound directly to Bombyx mori microtubule-associated protein 1-light chain 3 (BmLC3), an autophagosome marker, and was colocalized with BmLC3 to the isolation membrane of autophagosome, implying the involvement of polyhedrin in cellular autophagy. Inhibition of autophagy by 3-methyladenine (3-MA) dramatically resulted in decrease of polyhedrin expression and polyhedra particle production. These observations suggested that highly expressed polyhedrin forms aggregate to get involved in cellular autophagy then play an important role in polyhedra production.

No MeSH data available.


Related in: MedlinePlus