title: Proteomics of the 26S proteasome in Spodoptera frugiperda cells infected with the nucleopolyhedrovirus, AcMNPV.
abstract: Baculoviruses are large DNA viruses that infect insect species such as Lepidoptera and are used in biotechnology for protein production and in agriculture as insecticides against crop pests. Baculoviruses require activity of host proteasomes for efficient reproduction, but how they control the cellular proteome and interact with the ubiquitin proteasome system (UPS) of infected cells remains unknown. In this report, we analyzed possible changes in the subunit composition of 26S proteasomes of the fall armyworm, Spodoptera frugiperda (Sf9), cells in the course of infection with the Autographa californica multiple nucleopolyhedrovirus (AcMNPV). 26S proteasomes were purified from Sf9 cells by an immune affinity method and subjected to 2D gel electrophoresis followed by MALDI-TOF mass spectrometry and Mascot search in bioinformatics databases. A total of 34 homologues of 26S proteasome subunits of eukaryotic species were identified including 14 subunits of the 20S core particle (7 α and 7 β subunits) and 20 subunits of the 19S regulatory particle (RP). The RP contained homologues of 11 of RPN-type and 6 of RPT-type subunits, 2 deubiquitinating enzymes (UCH-14/UBP6 and UCH-L5/UCH37), and thioredoxin. Similar 2D-gel maps of 26S proteasomes purified from uninfected and AcMNPV-infected cells at 48hpi confirmed the structural integrity of the 26S proteasome in insect cells during baculovirus infection. However, subtle changes in minor forms of some proteasome subunits were detected. A portion of the α5(zeta) cellular pool that presumably was not associated with the proteasome underwent partial proteolysis at a late stage in infection.
authors.affiliation: N.K. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, 26 Vavilova Str., Moscow 119334, Russia.
authors.forename: Yulia V
authors.initials: YV
authors.lastname: Lyupina
authors.affiliation: V.A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119334, Russia.
authors.forename: Olga G
authors.initials: OG
authors.lastname: Zatsepina
authors.affiliation: A.N. Belozersky Institute of Physico-Chemical Biology MSU, 1c40 Leninskie Gory, Moscow 119234, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Serebryakova
authors.affiliation: N.K. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, 26 Vavilova Str., Moscow 119334, Russia.
authors.forename: Pavel A
authors.initials: PA
authors.lastname: Erokhov
authors.affiliation: N.K. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, 26 Vavilova Str., Moscow 119334, Russia.
authors.forename: Svetlana B
authors.initials: SB
authors.lastname: Abaturova
authors.affiliation: N.K. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, 26 Vavilova Str., Moscow 119334, Russia.
authors.forename: Oksana I
authors.initials: OI
authors.lastname: Kravchuk
authors.affiliation: V.A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119334, Russia.
authors.forename: Olga V
authors.initials: OV
authors.lastname: Orlova
authors.affiliation: V.A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119334, Russia.
authors.forename: Svetlana N
authors.initials: SN
authors.lastname: Beljelarskaya
authors.affiliation: M.V. Lomonosov Moscow State University, Faculty of Biology, 1-12 Leninskie Gory, Moscow 119234, Russia.
authors.forename: Andrey I
authors.initials: AI
authors.lastname: Lavrov
authors.affiliation: M.V. Lomonosov Moscow State University, Faculty of Biology, 1-12 Leninskie Gory, Moscow 119234, Russia.
authors.forename: Olga S
authors.initials: OS
authors.lastname: Sokolova
authors.affiliation: N.K. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, 26 Vavilova Str., Moscow 119334, Russia. Electronic address: mikhailov48@mail.ru.
authors.forename: Victor S
authors.initials: VS
authors.lastname: Mikhailov
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000472:pathogenicity*; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry*; D040901:Proteomics; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018411:Spodoptera / Q000166:cytology*
chemical_list: D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease
keywords: 20S proteasome; 26S proteasome; AcMNPV; Baculovirus; Nucleopolyhedrovirus; Spodoptera frugiperda