Abstract
The cell entry mechanism of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is not fully understood. Previous studies showed that AcMNPV entered host cells primarily through clathrin-mediated endocytosis, and could efficiently infect cells via fusion with the plasma membrane after a low-pH trigger. However, whether AcMNPV enters cells via these two pathways simultaneously, and the exact manner in which AcMNPV particles are internalized into cells remains unclear. In this study, using single-virus tracking, we observed that AcMNPV particles were first captured by pre-existing clathrin-coated pits (CCP), and were then delivered to early endosomes. Population-based analysis of single-virus tracking and quantitative electron microscopy demonstrated that the majority of particles were captured by CCPs and internalized via invagination. In contrast, a minority of virus particles were not delivered to CCPs, and were internalized through direct fusion with the plasma membrane without invagination. Quantitative electron microscopy also showed that, while inhibition of CCP assembly significantly impaired viral internalization, inhibition of endosomal acidification blocked virus particles out of vesicles. Collectively, these findings demonstrated that approximately 90% of AcMNPV particles entered cells through clathrin-mediated endocytosis and 10% entered via direct fusion with the plasma membrane. This study will lead toward a better understanding of AcMNPV infection.
Highlights
Baculoviruses, belonging to the baculoviridae, are enveloped DNA viruses that have been extensively applied for insect control and eukaryotic expression, and have potential applications in gene therapy [1,2,3,4,5,6]
The results showed that inhibition of the assembly of clathrin-coated pits (CCP) with CPZ significantly inhibited the internalization of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) virus particles compared with control cells, indicating that CCPs are required for viral internalization (Figure 2E)
Our findings demonstrated that AcMNPV particles primarily enter cells through clathrin-dependent endocytosis, with a small fraction entering through direct fusion with the plasma membrane
Summary
Baculoviruses, belonging to the baculoviridae, are enveloped DNA viruses that have been extensively applied for insect control and eukaryotic expression, and have potential applications in gene therapy [1,2,3,4,5,6]. Autographa californica multiple nucleopolyhedrovirus (AcMNPV), a model virus of the baculoviridae, has been extensively studied [8,9,10,11,12,13]. Further development of these applications requires further insight into the cell entry pathway. We investigated the cell entry pathway of the BV of AcMNPV
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