Abstract
Baculovirus infection of Spodoptera frugiperda cells is a system of choice to produce a range of recombinant proteins, vaccines and, potentially, gene therapy vectors. While baculovirus genomes are well characterized, the genome of S. frugiperda is not sequenced and the virus-host molecular interplay is sparsely known. Herein, we describe the application of stable isotope labeling by amino acids in cell culture (SILAC) to obtain the first comparative proteome quantitation of S. frugiperda cells during growth and early baculovirus infection. The proteome coverage was maximized by compiling a search database with protein annotations from insect species. Of interest were differentially proteins related to energy metabolism, endoplasmic reticulum and oxidative stress, yet not investigated in the scope of baculovirus infection. Further, the reduced expression of key viral-encoded proteins early in the infection cycle is suggested to be related with decreased viral replication at high cell density culture. These findings have implications for virological research and improvement of baculovirus-based bioprocesses.
Highlights
Baculoviruses (BVs) are a diverse family of arthropod pathogens containing large DNA genomes reaching up to 180 kbp [1]
This missing knowledge was overcome by compiling a search database with available annotations from related insect species, constructed as a subset of the non–redundant protein database maintained by the National Center for Biotechnology Information (NCBI)
To provide an estimation of interspecies redundancy, the Protein Information Resource SuperFamily (PIRSF) classification system was adopted to group proteins sharing homology and homeomorphy [21]. 233 out of 418 proteins could be assigned to unique PIRSF designations, indicating a total of 56% different Sf9 proteins
Summary
Baculoviruses (BVs) are a diverse family of arthropod pathogens containing large DNA genomes reaching up to 180 kbp [1]. BVs vary considerably in their genome composition and in general have a narrow host range; the prototype Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) infects a few insect lepidopteran species. The economic interest of BVs derives from their use as biopesticides and, along the past decades, as versatile protein expression systems [2]. Due to their inability to replicate in mammalian cells, their use as antigen displaying vectors and gene delivery vehicles has become exceedingly popular in recent years [3]. The host for AcMNPV propagation is usually a cell line derived from the fall armyworm S. frugiperda, the Sf9 clonal isolate. More than a vehicle for viral propagation, insect cell lines are chosen for their robust growth, straightforward culture scale-up and post-translational modifications of expressed proteins [4]
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