Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i>
To understand the mechanism of replication used by baculoviruses, it is essential to describe all the factors involved, including virus and host proteins and the sequences where DNA synthesis starts. A lot of work on this topic has been done, but there is still confusion in defining what sequence/s...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2019-07-01
|
Series: | Viruses |
Subjects: | |
Online Access: | https://www.mdpi.com/1999-4915/11/7/648 |
id |
doaj-76bdde2bc88f4148893d0e0a1dbaba65 |
---|---|
record_format |
Article |
spelling |
doaj-76bdde2bc88f4148893d0e0a1dbaba652020-11-24T21:30:44ZengMDPI AGViruses1999-49152019-07-0111764810.3390/v11070648v11070648Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i>Solange A.B. Miele0Carolina S. Cerrudo1Cintia N. Parsza2María Victoria Nugnes3Diego L. Mengual Gómez4Mariano N. Belaich5P. Daniel Ghiringhelli6Laboratorio de Ingeniería Genética y Biología Celular y Molecular-Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada (IMBA), Universidad Nacional de Quilmes, CONICET, Bernal B1876BXD, ArgentinaLaboratorio de Ingeniería Genética y Biología Celular y Molecular-Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada (IMBA), Universidad Nacional de Quilmes, CONICET, Bernal B1876BXD, ArgentinaLaboratorio de Ingeniería Genética y Biología Celular y Molecular-Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada (IMBA), Universidad Nacional de Quilmes, CONICET, Bernal B1876BXD, ArgentinaLaboratorio de Ingeniería Genética y Biología Celular y Molecular-Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada (IMBA), Universidad Nacional de Quilmes, CONICET, Bernal B1876BXD, ArgentinaLaboratorio de Oncología Molecular, Universidad Nacional de Quilmes, CONICET, Bernal B1876BXD, ArgentinaLaboratorio de Ingeniería Genética y Biología Celular y Molecular-Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada (IMBA), Universidad Nacional de Quilmes, CONICET, Bernal B1876BXD, ArgentinaLaboratorio de Ingeniería Genética y Biología Celular y Molecular-Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada (IMBA), Universidad Nacional de Quilmes, CONICET, Bernal B1876BXD, ArgentinaTo understand the mechanism of replication used by baculoviruses, it is essential to describe all the factors involved, including virus and host proteins and the sequences where DNA synthesis starts. A lot of work on this topic has been done, but there is still confusion in defining what sequence/s act in such functions, and the mechanism of replication is not very well understood. In this work, we performed an AgMNPV replication kinetics into the susceptible UFL-Ag-286 cells to estimate viral genome synthesis rates. We found that the viral DNA exponentially increases in two different phases that are temporally separated by an interval of 5 h, probably suggesting the occurrence of two different mechanisms of replication. Then, we prepared a plasmid library containing virus fragments (0.5−2 kbp), which were transfected and infected with AgMNPV in UFL-Ag-286 cells. We identified 12 virus fragments which acted as origins of replication (ORI). Those fragments are in close proximity to core genes. This association to the core genome would ensure vertical transmission of ORIs. We also predict the presence of common structures on those fragments that probably recruit the replication machinery, a structure also present in previously reported ORIs in baculoviruses.https://www.mdpi.com/1999-4915/11/7/648baculovirusAgMNPVreplication |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Solange A.B. Miele Carolina S. Cerrudo Cintia N. Parsza María Victoria Nugnes Diego L. Mengual Gómez Mariano N. Belaich P. Daniel Ghiringhelli |
spellingShingle |
Solange A.B. Miele Carolina S. Cerrudo Cintia N. Parsza María Victoria Nugnes Diego L. Mengual Gómez Mariano N. Belaich P. Daniel Ghiringhelli Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i> Viruses baculovirus AgMNPV replication |
author_facet |
Solange A.B. Miele Carolina S. Cerrudo Cintia N. Parsza María Victoria Nugnes Diego L. Mengual Gómez Mariano N. Belaich P. Daniel Ghiringhelli |
author_sort |
Solange A.B. Miele |
title |
Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i> |
title_short |
Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i> |
title_full |
Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i> |
title_fullStr |
Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i> |
title_full_unstemmed |
Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of <i>Anticarsia gemmatalis</i> |
title_sort |
identification of multiple replication stages and origins in the nucleopolyhedrovirus of <i>anticarsia gemmatalis</i> |
publisher |
MDPI AG |
series |
Viruses |
issn |
1999-4915 |
publishDate |
2019-07-01 |
description |
To understand the mechanism of replication used by baculoviruses, it is essential to describe all the factors involved, including virus and host proteins and the sequences where DNA synthesis starts. A lot of work on this topic has been done, but there is still confusion in defining what sequence/s act in such functions, and the mechanism of replication is not very well understood. In this work, we performed an AgMNPV replication kinetics into the susceptible UFL-Ag-286 cells to estimate viral genome synthesis rates. We found that the viral DNA exponentially increases in two different phases that are temporally separated by an interval of 5 h, probably suggesting the occurrence of two different mechanisms of replication. Then, we prepared a plasmid library containing virus fragments (0.5−2 kbp), which were transfected and infected with AgMNPV in UFL-Ag-286 cells. We identified 12 virus fragments which acted as origins of replication (ORI). Those fragments are in close proximity to core genes. This association to the core genome would ensure vertical transmission of ORIs. We also predict the presence of common structures on those fragments that probably recruit the replication machinery, a structure also present in previously reported ORIs in baculoviruses. |
topic |
baculovirus AgMNPV replication |
url |
https://www.mdpi.com/1999-4915/11/7/648 |
work_keys_str_mv |
AT solangeabmiele identificationofmultiplereplicationstagesandoriginsinthenucleopolyhedrovirusofianticarsiagemmatalisi AT carolinascerrudo identificationofmultiplereplicationstagesandoriginsinthenucleopolyhedrovirusofianticarsiagemmatalisi AT cintianparsza identificationofmultiplereplicationstagesandoriginsinthenucleopolyhedrovirusofianticarsiagemmatalisi AT mariavictorianugnes identificationofmultiplereplicationstagesandoriginsinthenucleopolyhedrovirusofianticarsiagemmatalisi AT diegolmengualgomez identificationofmultiplereplicationstagesandoriginsinthenucleopolyhedrovirusofianticarsiagemmatalisi AT marianonbelaich identificationofmultiplereplicationstagesandoriginsinthenucleopolyhedrovirusofianticarsiagemmatalisi AT pdanielghiringhelli identificationofmultiplereplicationstagesandoriginsinthenucleopolyhedrovirusofianticarsiagemmatalisi |
_version_ |
1725961952705904640 |