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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 mediates HSPD1 protein stability.(A) Effect of dsATAD3A mediated knockdown of BmATAD3A on HSPD1 transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsATAD3A (1 μg), dsATAD3A (2 μg) and dsATAD3A (4 μg). After 48 h.p.t., the transcript levels of HSPD1 were examined by RT-PCR. (B) After 48 h.p.t., the expression levels of HSPD1 were examined by Western blotting. Tubulin expression levels as control was assessed. (C) Effect of dsHSPD1 mediated knockdown of BmHSPD1 on ATAD3A transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsHSPD1 (1 μg), dsHSPD1 (2 μg) and dsHSPD1 (4 μg). After 48 h.p.t., the transcript levels of ATAD3A were examined by RT-PCR. (D) After 48 h.p.t., the expression levels of ATAD3A were examined by Western blotting. Tubulin expression levels as control was assessed.
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f6: ATAD3A mediates HSPD1 protein stability.(A) Effect of dsATAD3A mediated knockdown of BmATAD3A on HSPD1 transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsATAD3A (1 μg), dsATAD3A (2 μg) and dsATAD3A (4 μg). After 48 h.p.t., the transcript levels of HSPD1 were examined by RT-PCR. (B) After 48 h.p.t., the expression levels of HSPD1 were examined by Western blotting. Tubulin expression levels as control was assessed. (C) Effect of dsHSPD1 mediated knockdown of BmHSPD1 on ATAD3A transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsHSPD1 (1 μg), dsHSPD1 (2 μg) and dsHSPD1 (4 μg). After 48 h.p.t., the transcript levels of ATAD3A were examined by RT-PCR. (D) After 48 h.p.t., the expression levels of ATAD3A were examined by Western blotting. Tubulin expression levels as control was assessed.

Mentions: To determine whether ATAD3A had a role in HSPD1 function, we used RT-PCR and Western blotting to analyze the knockdown of the ATAD3A and HSPD1 in BmN-SWU1 cells, respectively. Results showed that ATAD3A transcription levels and protein levels had no obvious changes after silencing HSPD1 with the dsHSPD1 compared with the control dsEGFP (Fig. 6A,B). These results indicated that HSPD1 has no significant effect on the expression of ATAD3A protein. In order to further confirm the regulatory relationship between ATAD3A and HSPD1, HSPD1 transcription levels and protein levels were detected in dsATAD3A transfected cells. Results showed that HSPD1 transcription levels and protein levels were reduced in dsATAD3A transfected cells compared with the control dsEGFP transfected cells (Fig. 6C,D). These results indicated that ATAD3A was important for the expression of HSPD1 protein. Combined with the results of their co-localization, we speculated that ATAD3A is an important regulator of HSPD1 protein expression and ATAD3A may be an upstream regulatory protein of HSPD1.


Baculovirus LEF-11 Hijack Host ATPase ATAD3A to Promote Virus Multiplication in Bombyx mori cells
ATAD3A mediates HSPD1 protein stability.(A) Effect of dsATAD3A mediated knockdown of BmATAD3A on HSPD1 transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsATAD3A (1 μg), dsATAD3A (2 μg) and dsATAD3A (4 μg). After 48 h.p.t., the transcript levels of HSPD1 were examined by RT-PCR. (B) After 48 h.p.t., the expression levels of HSPD1 were examined by Western blotting. Tubulin expression levels as control was assessed. (C) Effect of dsHSPD1 mediated knockdown of BmHSPD1 on ATAD3A transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsHSPD1 (1 μg), dsHSPD1 (2 μg) and dsHSPD1 (4 μg). After 48 h.p.t., the transcript levels of ATAD3A were examined by RT-PCR. (D) After 48 h.p.t., the expression levels of ATAD3A were examined by Western blotting. Tubulin expression levels as control was assessed.
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f6: ATAD3A mediates HSPD1 protein stability.(A) Effect of dsATAD3A mediated knockdown of BmATAD3A on HSPD1 transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsATAD3A (1 μg), dsATAD3A (2 μg) and dsATAD3A (4 μg). After 48 h.p.t., the transcript levels of HSPD1 were examined by RT-PCR. (B) After 48 h.p.t., the expression levels of HSPD1 were examined by Western blotting. Tubulin expression levels as control was assessed. (C) Effect of dsHSPD1 mediated knockdown of BmHSPD1 on ATAD3A transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsHSPD1 (1 μg), dsHSPD1 (2 μg) and dsHSPD1 (4 μg). After 48 h.p.t., the transcript levels of ATAD3A were examined by RT-PCR. (D) After 48 h.p.t., the expression levels of ATAD3A were examined by Western blotting. Tubulin expression levels as control was assessed.
Mentions: To determine whether ATAD3A had a role in HSPD1 function, we used RT-PCR and Western blotting to analyze the knockdown of the ATAD3A and HSPD1 in BmN-SWU1 cells, respectively. Results showed that ATAD3A transcription levels and protein levels had no obvious changes after silencing HSPD1 with the dsHSPD1 compared with the control dsEGFP (Fig. 6A,B). These results indicated that HSPD1 has no significant effect on the expression of ATAD3A protein. In order to further confirm the regulatory relationship between ATAD3A and HSPD1, HSPD1 transcription levels and protein levels were detected in dsATAD3A transfected cells. Results showed that HSPD1 transcription levels and protein levels were reduced in dsATAD3A transfected cells compared with the control dsEGFP transfected cells (Fig. 6C,D). These results indicated that ATAD3A was important for the expression of HSPD1 protein. Combined with the results of their co-localization, we speculated that ATAD3A is an important regulator of HSPD1 protein expression and ATAD3A may be an upstream regulatory protein of HSPD1.

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.