Autographa californica multiple nucleopolyhedrovirus ac75 is required for egress of nucleocapsids from the nucleus and formation of de novo intranuclear membrane microvesicles

Autor: Shi-Hui Fu, Lu-Lin Li, Ya-Jun Guo
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Genes
Viral
viruses
Gene Expression
lcsh:Medicine
Virus Replication
Nucleocapsids
Biochemistry
Fluorescence Microscopy
Virogenic stroma
Sf9 Cells
lcsh:Science
Microscopy
Microscopy
Confocal
Multidisciplinary
biology
Chemistry
Light Microscopy
Cell biology
Nucleic acids
medicine.anatomical_structure
Viral Envelope
Gene Knockdown Techniques
Cellular Structures and Organelles
Research Article
Plasmids
Nuclear Envelope
Blotting
Western
Viral Structure
DNA replication
Transfection
Research and Analysis Methods
Real-Time Polymerase Chain Reaction
Microbiology
03 medical and health sciences
Nuclear Membrane
Microscopy
Electron
Transmission
Viral envelope
Virology
Genetics
medicine
Animals
Nuclear membrane
Molecular Biology Techniques
Nucleocapsid
Molecular Biology
Cell Nucleus
fungi
lcsh:R
Biology and Life Sciences
Biological Transport
Cell Biology
DNA
biology.organism_classification
Viral Replication
Nucleopolyhedroviruses
Microvesicles
Autographa californica
030104 developmental biology
Viral replication
Cytoplasm
DNA
Viral
lcsh:Q
Nucleus
Zdroj: PLoS ONE, Vol 12, Iss 10, p e0185630 (2017)
PLoS ONE
ISSN: 1932-6203
Popis: In this study, Autographa californica multiple nucleopolyhedrovirus ac75 was functionally characterized. Ac75 has homologs in all sequenced genomes of alphabaculoviruses, betabaculoviruses, and gammabaculoviruses. It was determined to encode a protein that is associated with the nucleocapsid of budded virus and with both envelope and nucleocapsids of occlusion-derived virus. Sf9 cells transfected by an ac75-knockout bacmid resulted in the infection being restricted to single cells. No budded virus were detected although viral DNA replication and late gene expression were unaffected. Electron microscopy revealed that the virogenic stroma, nucleocapsids and occlusion bodies appeared normal in the cells transfected by an ac75-knockout bacmid. However, the nucleocapsids were unenveloped, the occlusion bodies did not contain any virions or nucleocapsids, and no nucleocapsids were found outside the nucleus or spanning the nuclear membrane. In addition, de novo intranuclear membrane microvesicles that are the precursor of occlusion-derived virus envelopes were absent in the nuclei of transfected cells. Confocal microscopy showed that AC75 protein appeared in the cytoplasm as early as 6 hours post infection. It localized to the ring zone at the periphery of the nucleus from 15 to 24 hours post infection and demonstrated light blocky cloud-like distribution in the center of the nucleus. AC75 was found to co-immunoprecipitate with BV and ODV associated envelope protein ODV-E25. The data from this study suggest that ac75 is essential for induction of the intranuclear membrane microvesicles, it appears to be required for the intranuclear envelopment of nucleocapsids, and is also essential for egress of nucleocapsids from the nuclei, in infected cells.
Databáze: OpenAIRE
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