Limits...
Egress of HSV-1 capsid requires the interaction of VP26 and a cellular tetraspanin membrane protein.

Wang L, Liu L, Che Y, Wang L, Jiang L, Dong C, Zhang Y, Li Q - Virol. J. (2010)

Bottom Line: In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro.The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication.Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute Of Medical Biology, Chinese Academy of Medicine Science, Peking Union Medical College, Kunming 650118, P. R. China.

ABSTRACT
HSV-1 viral capsid maturation and egress from the nucleus constitutes a self-controlled process of interactions between host cytoplasmic membrane proteins and viral capsid proteins. In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro. The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication. Additionally, analysis of a recombinant virus HSV-1-UG showed that mutating VP26 resulted in a decreased viral replication rate and in aggregation of viral mutant capsids in the nucleus. Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

Show MeSH

Related in: MedlinePlus

VP26 mutant virus shows a delayed proliferation. The recombinant virus HSV1-UG with a GFP-UL35 fusion gene was used to infect Vero or KMB-17 cells at 1 MOI. Punctated fluorescence spots were observed in the nucleus during infection, which is similar to the recombinant HSV1 with GFP-Vp16 fusion gene ([31]). At 12, 22, 24, 27, 36, 46, 60 and 72 h post-infection, samples of cell infected by HSV1-UG were collected and measured by real-time PCR with specific primers against α-4 gene. n = 3 for all time points. Error bars represent the standard error of the mean. Meanwhile, the KMB-17 cells infected by HSV1-UG were fixed with 5% glutaraldehyde and observed under the electron microscope. a. The Vero cells infected by VP26 mutant HSV1-UG were observed under fluorescence microscope at 12, 16 and 24 h. b. Growth curve of HSV1-UG compared with that of wild type HSV-1 in Vero cells. c. Electro-microscope observation of cells infected by HSV1-UG(X30, 000).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2913958&req=5

Figure 7: VP26 mutant virus shows a delayed proliferation. The recombinant virus HSV1-UG with a GFP-UL35 fusion gene was used to infect Vero or KMB-17 cells at 1 MOI. Punctated fluorescence spots were observed in the nucleus during infection, which is similar to the recombinant HSV1 with GFP-Vp16 fusion gene ([31]). At 12, 22, 24, 27, 36, 46, 60 and 72 h post-infection, samples of cell infected by HSV1-UG were collected and measured by real-time PCR with specific primers against α-4 gene. n = 3 for all time points. Error bars represent the standard error of the mean. Meanwhile, the KMB-17 cells infected by HSV1-UG were fixed with 5% glutaraldehyde and observed under the electron microscope. a. The Vero cells infected by VP26 mutant HSV1-UG were observed under fluorescence microscope at 12, 16 and 24 h. b. Growth curve of HSV1-UG compared with that of wild type HSV-1 in Vero cells. c. Electro-microscope observation of cells infected by HSV1-UG(X30, 000).

Mentions: In order to further investigate the biological events in the viral enveloping process generated by the interactions of VP26 and cellular CTMP-7, we constructed a viral mutant, HSV-1-UG, with a GFP-coding sequence fused to the UL35 gene. In Vero cells infected with HSV-1-UG, a mutant VP26 was expressed and distributed in fluorescent punctate spots throughout the infected cells and maintained this pattern until 24 h post-infection (see Fig.7a). Similar observations were found in the mutant HSV-1 infected HeLa cells and neuroma SH-5YSY cells (data not shown). However, the kinetic growth rate of this viral mutant was significantly lower than that of wild-type HSV-1. Interestingly, when the HSV-1-UG mutant was used to infect normal human embryonic lung fibroblast KMB-17 cells, aggregation of viral capsids in the cells was observed (Fig.7b). These data support the previous work performed in our laboratory.


Egress of HSV-1 capsid requires the interaction of VP26 and a cellular tetraspanin membrane protein.

Wang L, Liu L, Che Y, Wang L, Jiang L, Dong C, Zhang Y, Li Q - Virol. J. (2010)

VP26 mutant virus shows a delayed proliferation. The recombinant virus HSV1-UG with a GFP-UL35 fusion gene was used to infect Vero or KMB-17 cells at 1 MOI. Punctated fluorescence spots were observed in the nucleus during infection, which is similar to the recombinant HSV1 with GFP-Vp16 fusion gene ([31]). At 12, 22, 24, 27, 36, 46, 60 and 72 h post-infection, samples of cell infected by HSV1-UG were collected and measured by real-time PCR with specific primers against α-4 gene. n = 3 for all time points. Error bars represent the standard error of the mean. Meanwhile, the KMB-17 cells infected by HSV1-UG were fixed with 5% glutaraldehyde and observed under the electron microscope. a. The Vero cells infected by VP26 mutant HSV1-UG were observed under fluorescence microscope at 12, 16 and 24 h. b. Growth curve of HSV1-UG compared with that of wild type HSV-1 in Vero cells. c. Electro-microscope observation of cells infected by HSV1-UG(X30, 000).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2913958&req=5

Figure 7: VP26 mutant virus shows a delayed proliferation. The recombinant virus HSV1-UG with a GFP-UL35 fusion gene was used to infect Vero or KMB-17 cells at 1 MOI. Punctated fluorescence spots were observed in the nucleus during infection, which is similar to the recombinant HSV1 with GFP-Vp16 fusion gene ([31]). At 12, 22, 24, 27, 36, 46, 60 and 72 h post-infection, samples of cell infected by HSV1-UG were collected and measured by real-time PCR with specific primers against α-4 gene. n = 3 for all time points. Error bars represent the standard error of the mean. Meanwhile, the KMB-17 cells infected by HSV1-UG were fixed with 5% glutaraldehyde and observed under the electron microscope. a. The Vero cells infected by VP26 mutant HSV1-UG were observed under fluorescence microscope at 12, 16 and 24 h. b. Growth curve of HSV1-UG compared with that of wild type HSV-1 in Vero cells. c. Electro-microscope observation of cells infected by HSV1-UG(X30, 000).
Mentions: In order to further investigate the biological events in the viral enveloping process generated by the interactions of VP26 and cellular CTMP-7, we constructed a viral mutant, HSV-1-UG, with a GFP-coding sequence fused to the UL35 gene. In Vero cells infected with HSV-1-UG, a mutant VP26 was expressed and distributed in fluorescent punctate spots throughout the infected cells and maintained this pattern until 24 h post-infection (see Fig.7a). Similar observations were found in the mutant HSV-1 infected HeLa cells and neuroma SH-5YSY cells (data not shown). However, the kinetic growth rate of this viral mutant was significantly lower than that of wild-type HSV-1. Interestingly, when the HSV-1-UG mutant was used to infect normal human embryonic lung fibroblast KMB-17 cells, aggregation of viral capsids in the cells was observed (Fig.7b). These data support the previous work performed in our laboratory.

Bottom Line: In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro.The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication.Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute Of Medical Biology, Chinese Academy of Medicine Science, Peking Union Medical College, Kunming 650118, P. R. China.

ABSTRACT
HSV-1 viral capsid maturation and egress from the nucleus constitutes a self-controlled process of interactions between host cytoplasmic membrane proteins and viral capsid proteins. In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro. The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication. Additionally, analysis of a recombinant virus HSV-1-UG showed that mutating VP26 resulted in a decreased viral replication rate and in aggregation of viral mutant capsids in the nucleus. Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

Show MeSH
Related in: MedlinePlus