Physical mapping of Malacosoma neustria nuclear polyhedrosis virus genome and its modification in Antheraea pernyi cell culture

Kikhno, I. M.; Strokovskaya, L. I.; Meleshko, R. A.; Michalic, J.; Solomko, A. P.

doi:10.7124/bc.000630

Cited by 3 publications

(4 citation statements)

References 9 publications

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“…The linearization site was previously delineated on the physical map of the ManeNPV genome with respect to the Bam HI, Kpn I, and Pst I cleavage sites [23] . In the present study, the Bam HI-D fragment of the genome harboring the linearization site was cloned into pBR325.…”

Section: Resultsmentioning

confidence: 99%

“…An examination of the nucleotide sequence of the ManeNPV Bgl II- Hind III fragment ( Figure S2 ) revealed that the Pst I site, which is located proximal to the linearization site on the ManeNPV physical map [23] , is situated 17 bp upstream of the CE. Based on the comparison of the Pst I site position (17 bp upstream of the CE, in accordance with the sequencing data, and less than 100 bp upstream of the linearization site, in accordance with the ManeNPV physical map, with the CE size (155 bp), it was concluded that the linearization site must be located within the CE.…”

Section: Resultsmentioning

confidence: 99%

“…The infection process characteristic of this virus-cell system was associated with the emergence of linear viral genomes, a phenomenon that has never been reported before in relation to other baculovirus-host systems. The ''break site'', referred to herein as the ''linearization site'', was localized to a defined locus on the ManeNPV genome map [23]. The terminal sequences of the linear ManeNPV genome were considered to be of great interest because, when linked together in a circular genome, the terminal sequences were expected to compose an nfe involved in some unexplored aspect of baculoviral genome activity that is associated with the conversion of the original circular DNA into a linear derivative.…”

Section: Introductionmentioning

confidence: 99%

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Identification of a Conserved Non-Protein-Coding Genomic Element that Plays an Essential Role in Alphabaculovirus Pathogenesis

Kikhno

2014

PLoS ONE

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Highly homologous sequences 154–157 bp in length grouped under the name of “conserved non-protein-coding element” (CNE) were revealed in all of the sequenced genomes of baculoviruses belonging to the genus Alphabaculovirus. A CNE alignment led to the detection of a set of highly conserved nucleotide clusters that occupy strictly conserved positions in the CNE sequence. The significant length of the CNE and conservation of both its length and cluster architecture were identified as a combination of characteristics that make this CNE different from known viral non-coding functional sequences. The essential role of the CNE in the Alphabaculovirus life cycle was demonstrated through the use of a CNE-knockout Autographa californica multiple nucleopolyhedrovirus (AcMNPV) bacmid. It was shown that the essential function of the CNE was not mediated by the presumed expression activities of the protein- and non-protein-coding genes that overlap the AcMNPV CNE. On the basis of the presented data, the AcMNPV CNE was categorized as a complex-structured, polyfunctional genomic element involved in an essential DNA transaction that is associated with an undefined function of the baculovirus genome.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of a Conserved Non-Protein-Coding Genomic Element that Plays an Essential Role in Alphabaculovirus Pathogenesis

Kikhno

2014

PLoS ONE

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“…The CNE was originally identified at the ends of the linear form of the Malacosoma neustria nucleopolyhedrovirus (ManeNPV) genome (45). The CNE has been demonstrated to act in cis for the BV production of AcMNPV and is suggested to be involved in the replication, recombination, or cleavage of the viral genome (41).…”

Section: Resultsmentioning

confidence: 99%

The Autographa californica Multiple Nucleopolyhedrovirus ac83 Gene Contains a cis -Acting Element That Is Essential for Nucleocapsid Assembly

Huang

Pan

Zhu

et al. 2017

J Virol

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Baculoviridae is a family of insect-specific viruses that have a circular double-stranded DNA genome packaged within a rod-shaped capsid. The mechanism of baculovirus nucleocapsid assembly remains unclear. Previous studies have shown that deletion of the ac83 gene of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) blocks viral nucleocapsid assembly. Interestingly, the ac83-encoded protein Ac83 is not a component of the nucleocapsid, implying a particular role for ac83 in nucleocapsid assembly that may be independent of its protein product. To examine this possibility, Ac83 synthesis was disrupted by insertion of a chloramphenicol resistance gene into its coding sequence or by deleting its promoter and translation start codon. Both mutants produced progeny viruses normally, indicating that the Ac83 protein is not required for nucleocapsid assembly. Subsequently, complementation assays showed that the production of progeny viruses required the presence of ac83 in the AcMNPV genome instead of its presence in trans. Therefore, we reasoned that ac83 is involved in nucleocapsid assembly via an internal cis-acting element, which we named the nucleocapsid assembly-essential element (NAE). The NAE was identified to lie within nucleotides 1651 to 1850 of ac83 and had 8 conserved A/T-rich regions. Sequences homologous to the NAE were found only in alphabaculoviruses and have a conserved positional relationship with another essential cis-acting element that was recently identified. The identification of the NAE may help to connect the data of viral cis-acting elements and related proteins in the baculovirus nucleocapsid assembly, which is important for elucidating DNA-protein interaction events during this process.IMPORTANCE Virus nucleocapsid assembly usually requires specific cis-acting elements in the viral genome for various processes, such as the selection of the viral genome from the cellular nucleic acids, the cleavage of concatemeric viral genome replication intermediates, and the encapsidation of the viral genome into procapsids. In linear DNA viruses, such elements generally locate at the ends of the viral genome; however, most of these elements remain unidentified in circular DNA viruses (including baculovirus) due to their circular genomic conformation. Here, we identified a nucleocapsid assembly-essential element in the AcMNPV (the archetype of baculovirus) genome. This finding provides an important reference for studies of nucleocapsid assembly-related elements in baculoviruses and other circular DNA viruses. Moreover, as most of the previous studies of baculovirus nucleocapsid assembly have been focused on viral proteins, our study provides a novel entry point to investigate this mechanism via cis-acting elements in the viral genome.

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Complete genome sequence analysis of the Malacosoma neustria nucleopolyhedrovirus from Turkey

et al. 2018

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The lackey moth, Malacosoma neustria (Linnaeus, 1758), a worldwide pest, causes extensive economic losses particularly on hazelnut, plum, oak, poplar, and willow trees. A baculovirus, Malacosoma neustria nucleopolyhedrovirus (ManeNPV-T2), has been isolated from the larvae collected in Turkey and appears to have a potential as a microbial control agent. In this study, we describe the complete genome sequence of ManeNPV-T2 and compare it to other sequenced baculovirus genomes. The ManeNPV-T2 genome is a circular double-stranded DNA molecule of 130,202 bp, has 38.2% G + C, and is predicted to contain 131 putative open reading frames (ORFs) each with a coding capacity of more then 50 amino acids. There are 27 ORFs with unknown function of which 6 are unique to ManeNPV-T2. Eleven homologous regions (hrs) and two bro genes (bro-a and bro-b) were identified in the genome. There are two homologues of chaB and nicotinamide riboside kinase-1 genes, separated from themselves with a few nucleotides. Additionally, ac145, thought to be per os infectivity factor (pif) gene, is also found as two homologues. All 38 core genes are found in the ManeNPV-T2 genome. The phylogenetic tree of ManeNPV-T2 in relation to 50 other baculoviruses whose genomes have been completely sequenced showed ManeNPV-T2 to be closely related to the group II NPVs. This study expands our knowledge on baculoviruses, describes the characterization ManeNPV, and ultimately contributes to the registration of this virus as a microbial pesticide.

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Physical mapping of Malacosoma neustria nuclear polyhedrosis virus genome and its modification in Antheraea pernyi cell culture

Cited by 3 publications

References 9 publications

Identification of a Conserved Non-Protein-Coding Genomic Element that Plays an Essential Role in Alphabaculovirus Pathogenesis

Identification of a Conserved Non-Protein-Coding Genomic Element that Plays an Essential Role in Alphabaculovirus Pathogenesis

The Autographa californica Multiple Nucleopolyhedrovirus ac83 Gene Contains a cis -Acting Element That Is Essential for Nucleocapsid Assembly

Complete genome sequence analysis of the Malacosoma neustria nucleopolyhedrovirus from Turkey

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