Limits...
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.


LEF-11 induces the expression of ATAD3A and HSPD1.(A) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in LEF-11 transfected cells. At the indicated time points, cells were harvested and total RNA was prepared and reverse transcription reactions were performed with SYBR Select Master Mix Reagent. (B) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in WT and KO transfected cells. Error bars indicate standard deviations from the mean. NS, not significant; **represents statistically significant differences at the level of P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5385504&req=5

f2: LEF-11 induces the expression of ATAD3A and HSPD1.(A) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in LEF-11 transfected cells. At the indicated time points, cells were harvested and total RNA was prepared and reverse transcription reactions were performed with SYBR Select Master Mix Reagent. (B) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in WT and KO transfected cells. Error bars indicate standard deviations from the mean. NS, not significant; **represents statistically significant differences at the level of P < 0.01.

Mentions: To further illustrate the relationship between BmNPV LEF-11 with ATAD3A and HSPD1 of the Bombyx mori in virus multiplication, we examined the effect of LEF-11 only on the expression of BmATAD3A and BmHSPD1 gene. BmN-SWU1 cells were transfected with lef-11 gene expression plasmids. At various time–points following transfection, cells were harvested and total RNA was prepared and subjected to qRT-PCR analysis with the indicated primers. At 0 h.p.t., a similar amount of BmATAD3A or BmHSPD1 transcription was detected in cells transfected with lef-11 gene expression plasmids and control cells transfected with pIZ-V5/His plasmids cells. As shown in Fig. 2ABmATAD3A or BmHSPD1 transcription levels significantly increased in lef-11 gene expression plasmids transfected cells. BmATAD3A transcription level was 9.2, 3.3, 4.5 and 4.1-fold greater in LEF-11 overexpressed cells than control pIZ-V5/His transfected cells at 12, 24, 48 and 72 h.p.t., respectively (Fig. 2A). In addition, BmHSPD1 transcription level increased 1.9, 2.3, 3.2 and 1.6-fold in lef-11 gene expression plasmids transfected cells compared with control pIZ-V5/His transfected cells at 12, 24, 48 and 72 h.p.t., respectively (Fig. 2A). These results indicated that BmNPV lef-11 gene expression could induce the levels of transcription of BmATAD3A and BmHSPD1.


Baculovirus LEF-11 Hijack Host ATPase ATAD3A to Promote Virus Multiplication in Bombyx mori cells
LEF-11 induces the expression of ATAD3A and HSPD1.(A) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in LEF-11 transfected cells. At the indicated time points, cells were harvested and total RNA was prepared and reverse transcription reactions were performed with SYBR Select Master Mix Reagent. (B) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in WT and KO transfected cells. Error bars indicate standard deviations from the mean. NS, not significant; **represents statistically significant differences at the level of P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5385504&req=5

f2: LEF-11 induces the expression of ATAD3A and HSPD1.(A) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in LEF-11 transfected cells. At the indicated time points, cells were harvested and total RNA was prepared and reverse transcription reactions were performed with SYBR Select Master Mix Reagent. (B) RT-PCR analysis of BmATAD3A or BmHSPD1 transcription in WT and KO transfected cells. Error bars indicate standard deviations from the mean. NS, not significant; **represents statistically significant differences at the level of P < 0.01.
Mentions: To further illustrate the relationship between BmNPV LEF-11 with ATAD3A and HSPD1 of the Bombyx mori in virus multiplication, we examined the effect of LEF-11 only on the expression of BmATAD3A and BmHSPD1 gene. BmN-SWU1 cells were transfected with lef-11 gene expression plasmids. At various time–points following transfection, cells were harvested and total RNA was prepared and subjected to qRT-PCR analysis with the indicated primers. At 0 h.p.t., a similar amount of BmATAD3A or BmHSPD1 transcription was detected in cells transfected with lef-11 gene expression plasmids and control cells transfected with pIZ-V5/His plasmids cells. As shown in Fig. 2ABmATAD3A or BmHSPD1 transcription levels significantly increased in lef-11 gene expression plasmids transfected cells. BmATAD3A transcription level was 9.2, 3.3, 4.5 and 4.1-fold greater in LEF-11 overexpressed cells than control pIZ-V5/His transfected cells at 12, 24, 48 and 72 h.p.t., respectively (Fig. 2A). In addition, BmHSPD1 transcription level increased 1.9, 2.3, 3.2 and 1.6-fold in lef-11 gene expression plasmids transfected cells compared with control pIZ-V5/His transfected cells at 12, 24, 48 and 72 h.p.t., respectively (Fig. 2A). These results indicated that BmNPV lef-11 gene expression could induce the levels of transcription of BmATAD3A and BmHSPD1.

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.