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Baculovirus LEF-11 Hijack Host ATPase ATAD3A to Promote Virus Multiplication in Bombyx mori cells

View Article: PubMed Central - PubMed

ABSTRACT

Research on molecular mechanisms that viruses use to regulate the host apparatus is important in virus infection control and antiviral therapy exploration. Our previous research showed that the Bombyx mori nucleopolyhedrovirus (BmNPV) LEF-11 localized to dense regions of the cell nucleus and is required for viral DNA replication. Herein, we examined the mechanism of LEF-11 on BmNPV multiplication and demonstrated that baculovirus LEF-11 interacts with Bombyx mori ATAD3A and HSPD1 (HSP60) protein. Furthermore, we showed that LEF-11 has the ability to induce and up-regulate the expression of ATAD3A and HSPD1, phenomena that were both reversed upon knockdown of lef-11. Our findings showed that ATAD3A and HSPD1 were necessary and contributed to BmNPV multiplication in Bombyx mori cells. Moreover, ATAD3A was found to directly interact with HSPD1. Interestingly, ATAD3A was required for the expression of HSPD1, while the knockdown of HSPD1 had no obvious effect on the expression level of ATAD3A. Taken together, the data presented in the current study demonstrated that baculovirus LEF-11 hijacks the host ATPase family members, ATAD3A and HSPD1, efficiently promote the multiplication of the virus. This study furthers our understanding of how baculovirus modulates energy metabolism of the host and provides a new insight into the molecular mechanisms of antiviral research.

No MeSH data available.


ATAD3A interacts with HSPD1.(A) Subcellular localization of ATAD3A and HSPD in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with FITC-labeled and Hoechst33258 at 48 h post-transfection in BmN-SWU1 cells. Green fluorescence represents fluorescent represent ATAD3A and HSPD1, blue fluorescence represents the nucleus. Scale bar: 5 μm. (B) Mitochondria co-location of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with Alexa 555-labeled, mitochondria-tracker and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents ATAD3A and HSPD1, Green fluorescence represents Mito-Tracker, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (C) Co-localization of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 stained with Alexa 555-labeled anti-HSPD1, FITC-labeled anti-ATAD3A and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents HSPD1, Green fluorescence represents ATAD3A, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (D) Co-immunoprecipitation of ATAD3A and HSPD1 examined by Western blotting. The label on the top of each panel shows the antibodies used for immunoprecipitation. The labels on the right of each panel shows the antibodies used for analysis of Western blotting. The apparent molecular size of each band is shown on the left of each panel.
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f5: ATAD3A interacts with HSPD1.(A) Subcellular localization of ATAD3A and HSPD in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with FITC-labeled and Hoechst33258 at 48 h post-transfection in BmN-SWU1 cells. Green fluorescence represents fluorescent represent ATAD3A and HSPD1, blue fluorescence represents the nucleus. Scale bar: 5 μm. (B) Mitochondria co-location of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with Alexa 555-labeled, mitochondria-tracker and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents ATAD3A and HSPD1, Green fluorescence represents Mito-Tracker, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (C) Co-localization of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 stained with Alexa 555-labeled anti-HSPD1, FITC-labeled anti-ATAD3A and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents HSPD1, Green fluorescence represents ATAD3A, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (D) Co-immunoprecipitation of ATAD3A and HSPD1 examined by Western blotting. The label on the top of each panel shows the antibodies used for immunoprecipitation. The labels on the right of each panel shows the antibodies used for analysis of Western blotting. The apparent molecular size of each band is shown on the left of each panel.

Mentions: To further determine the function of ATAD3A and HSPD1 in silkworm cells, we used confocal microscopy to analyze their localization after transfection in BmN-SWU1 cells. Immunofluorescence assay showed that ATAD3A and HSPD1 were mainly distributed in the cytoplasm (Fig. 5A). Mitochondria co-localization assay showed that they co-localize with mitochondria as assessed by Mito-Tracker Green (Fig. 5B). In contrast, the control DsRed protein fluorescence signals were observed in the cell nuclei and cytoplasm. These results further suggest that the endogenous ATAD3A and HSPD1 are mitochondrial proteins in silkworm cells.


Baculovirus LEF-11 Hijack Host ATPase ATAD3A to Promote Virus Multiplication in Bombyx mori cells
ATAD3A interacts with HSPD1.(A) Subcellular localization of ATAD3A and HSPD in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with FITC-labeled and Hoechst33258 at 48 h post-transfection in BmN-SWU1 cells. Green fluorescence represents fluorescent represent ATAD3A and HSPD1, blue fluorescence represents the nucleus. Scale bar: 5 μm. (B) Mitochondria co-location of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with Alexa 555-labeled, mitochondria-tracker and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents ATAD3A and HSPD1, Green fluorescence represents Mito-Tracker, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (C) Co-localization of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 stained with Alexa 555-labeled anti-HSPD1, FITC-labeled anti-ATAD3A and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents HSPD1, Green fluorescence represents ATAD3A, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (D) Co-immunoprecipitation of ATAD3A and HSPD1 examined by Western blotting. The label on the top of each panel shows the antibodies used for immunoprecipitation. The labels on the right of each panel shows the antibodies used for analysis of Western blotting. The apparent molecular size of each band is shown on the left of each panel.
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f5: ATAD3A interacts with HSPD1.(A) Subcellular localization of ATAD3A and HSPD in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with FITC-labeled and Hoechst33258 at 48 h post-transfection in BmN-SWU1 cells. Green fluorescence represents fluorescent represent ATAD3A and HSPD1, blue fluorescence represents the nucleus. Scale bar: 5 μm. (B) Mitochondria co-location of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 were stained with Alexa 555-labeled, mitochondria-tracker and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents ATAD3A and HSPD1, Green fluorescence represents Mito-Tracker, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (C) Co-localization of ATAD3A and HSPD1 in BmN-SWU1 cells. ATAD3A and HSPD1 stained with Alexa 555-labeled anti-HSPD1, FITC-labeled anti-ATAD3A and Hoechst33258 at 48 h post-transfection in the BmN-SWU1 cells. Red fluorescence represents HSPD1, Green fluorescence represents ATAD3A, and blue fluorescence represents the nucleus. Scale bar: 5 μm. (D) Co-immunoprecipitation of ATAD3A and HSPD1 examined by Western blotting. The label on the top of each panel shows the antibodies used for immunoprecipitation. The labels on the right of each panel shows the antibodies used for analysis of Western blotting. The apparent molecular size of each band is shown on the left of each panel.
Mentions: To further determine the function of ATAD3A and HSPD1 in silkworm cells, we used confocal microscopy to analyze their localization after transfection in BmN-SWU1 cells. Immunofluorescence assay showed that ATAD3A and HSPD1 were mainly distributed in the cytoplasm (Fig. 5A). Mitochondria co-localization assay showed that they co-localize with mitochondria as assessed by Mito-Tracker Green (Fig. 5B). In contrast, the control DsRed protein fluorescence signals were observed in the cell nuclei and cytoplasm. These results further suggest that the endogenous ATAD3A and HSPD1 are mitochondrial proteins in silkworm cells.

View Article: PubMed Central - PubMed

ABSTRACT

Research on molecular mechanisms that viruses use to regulate the host apparatus is important in virus infection control and antiviral therapy exploration. Our previous research showed that the Bombyx mori nucleopolyhedrovirus (BmNPV) LEF-11 localized to dense regions of the cell nucleus and is required for viral DNA replication. Herein, we examined the mechanism of LEF-11 on BmNPV multiplication and demonstrated that baculovirus LEF-11 interacts with Bombyx mori ATAD3A and HSPD1 (HSP60) protein. Furthermore, we showed that LEF-11 has the ability to induce and up-regulate the expression of ATAD3A and HSPD1, phenomena that were both reversed upon knockdown of lef-11. Our findings showed that ATAD3A and HSPD1 were necessary and contributed to BmNPV multiplication in Bombyx mori cells. Moreover, ATAD3A was found to directly interact with HSPD1. Interestingly, ATAD3A was required for the expression of HSPD1, while the knockdown of HSPD1 had no obvious effect on the expression level of ATAD3A. Taken together, the data presented in the current study demonstrated that baculovirus LEF-11 hijacks the host ATPase family members, ATAD3A and HSPD1, efficiently promote the multiplication of the virus. This study furthers our understanding of how baculovirus modulates energy metabolism of the host and provides a new insight into the molecular mechanisms of antiviral research.

No MeSH data available.