Changes in the Proteome of Langat-Infected Ixodes scapularis ISE6 Cells: Metabolic Pathways Associated with Flavivirus Infection

被引:39
作者
Grabowski, Jeffrey M. [1 ,2 ]
Perera, Rushika [2 ,4 ]
Roumani, Ali M. [3 ,5 ]
Hedrick, Victoria E. [3 ]
Inerowicz, Halina D. [3 ,6 ]
Hill, Catherine A. [1 ]
Kuhn, Richard J. [2 ,3 ]
机构
[1] Purdue Univ, Coll Agr, Dept Entomol, W Lafayette, IN 47907 USA
[2] Purdue Univ, Coll Sci, Dept Biol Sci, Markey Ctr Struct Biol, W Lafayette, IN 47907 USA
[3] Purdue Univ, Bindley Biosci Ctr, W Lafayette, IN 47907 USA
[4] Colorado State Univ, Dept Microbiol Immunol & Pathol, Arthropod Borne & Infect Dis Lab, Ft Collins, CO 80523 USA
[5] Gulf Univ Sci & Technol, Coll Arts & Sci, Dept Comp Sci, Mishref, Kuwait
[6] Purdue Univ, Off Indiana State Chem, W Lafayette, IN 47907 USA
关键词
BORNE ENCEPHALITIS-VIRUS; MASS-SPECTROMETRY; GLUTAMINE-METABOLISM; DOPAMINE-RECEPTORS; ALPHA-SUBUNIT; TICK; REPLICATION; DISCOVERY; EXPRESSION; PROTEINS;
D O I
10.1371/journal.pntd.0004180
中图分类号
R51 [传染病];
学科分类号
100201 [内科学];
摘要
Background Ticks (Family Ixodidae) transmit a variety of disease causing agents to humans and animals. The tick-borne flaviviruses (TBFs; family Flaviviridae) are a complex of viruses, many of which cause encephalitis and hemorrhagic fever, and represent global threats to human health and biosecurity. Pathogenesis has been well studied in human and animal disease models. Equivalent analyses of tick-flavivirus interactions are limited and represent an area of study that could reveal novel approaches for TBF control. Methodology/Principal Findings High resolution LC-MS/MS was used to analyze the proteome of Ixodes scapularis (Lyme disease tick) embryonic ISE6 cells following infection with Langat virus (LGTV) and identify proteins associated with viral infection and replication. Maximal LGTV infection of cells and determination of peak release of infectious virus, was observed at 36 hours post infection (hpi). Proteins were extracted from ISE6 cells treated with LGTV and non-infectious (UV inactivated) LGTV at 36 hpi and analyzed by mass spectrometry. The Omics Discovery Pipeline (ODP) identified thousands of MS peaks. Protein homology searches against the I. scapularis IscaW1 genome assembly identified a total of 486 proteins that were subsequently assigned to putative functional pathways using searches against the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. 266 proteins were differentially expressed following LGTV infection relative to non-infected (mock) cells. Of these, 68 proteins exhibited increased expression and 198 proteins had decreased expression. The majority of the former were classified in the KEGG pathways: "translation", "amino acid metabolism", and "protein folding/sorting/degradation". Finally, Trichostatin A and Oligomycin A increased and decreased LGTV replication in vitro in ISE6 cells, respectively. Conclusions/Significance Proteomic analyses revealed ISE6 proteins that were differentially expressed at the peak of LGTV replication. Proteins with increased expression following infection were associated with cellular metabolic pathways and glutaminolysis. In vitro assays using small molecules implicate malate dehydrogenase (MDH2), the citrate cycle, cellular acetylation, and electron transport chain processes in viral replication. Proteins were identified that may be required for TBF infection of ISE6 cells. These proteins are candidates for functional studies and targets for the development of transmission-blocking vaccines and drugs.
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页数:25
相关论文
共 80 条
[1]
Bridging membrane and cytoskeleton dynamics in the secretory and endocytic pathways [J].
Anitei, Mihaela ;
Hoflack, Bernard .
NATURE CELL BIOLOGY, 2012, 14 (01) :11-19
[2]
[Anonymous], CURR PROTOC IMMUNOL, DOI DOI 10.1002/0471142735IMA03BS21
[3]
Human immunodeficiency virus-1 Tat protein interacts with distinct proteasomal α and β subunits [J].
Apcher, GS ;
Heink, S ;
Zantopf, D ;
Kloetzel, PM ;
Schmid, HP ;
Mayer, RJ ;
Krüger, E .
FEBS LETTERS, 2003, 553 (1-2) :200-204
[4]
Global Cross-Talk of Genes of the Mosquito Aedes aegypti in Response to Dengue Virus Infection [J].
Behura, Susanta K. ;
Gomez-Machorro, Consuelo ;
Harker, Brent W. ;
Debruyn, Becky ;
Lovin, Diane D. ;
Hemme, Ryan R. ;
Mori, Akio ;
Romero-Severson, Jeanne ;
Severson, David W. .
PLOS NEGLECTED TROPICAL DISEASES, 2011, 5 (11)
[5]
Tick cell lines: tools for tick and tick-borne disease research [J].
Bell-Sakyi, Lesley ;
Zweygarth, Erich ;
Blouin, Edmour F. ;
Gould, Ernest A. ;
Jongejan, Frans .
TRENDS IN PARASITOLOGY, 2007, 23 (09) :450-457
[6]
Tick Cell Lines for Study of Crimean-Congo Hemorrhagic Fever Virus and Other Arboviruses [J].
Bell-Sakyi, Lesley ;
Kohl, Alain ;
Bente, Dennis A. ;
Fazakerley, John K. .
VECTOR-BORNE AND ZOONOTIC DISEASES, 2012, 12 (09) :769-781
[7]
CONTROLLING THE FALSE DISCOVERY RATE - A PRACTICAL AND POWERFUL APPROACH TO MULTIPLE TESTING [J].
BENJAMINI, Y ;
HOCHBERG, Y .
JOURNAL OF THE ROYAL STATISTICAL SOCIETY SERIES B-STATISTICAL METHODOLOGY, 1995, 57 (01) :289-300
[8]
Feeding Uninvited Guests: mTOR and AMPK Set the Table for Intracellular Pathogens [J].
Brunton, Jason ;
Steele, Shaun ;
Ziehr, Benjamin ;
Moorman, Nathaniel ;
Kawula, Thomas .
PLOS PATHOGENS, 2013, 9 (10)
[9]
Infectious cDNA clones of Langat tick-borne flavivirus that differ from their parent in peripheral neurovirulence [J].
Campbell, MS ;
Pletnev, AG .
VIROLOGY, 2000, 269 (01) :225-237
[10]
Glutamine Metabolism Is Essential for Human Cytomegalovirus Infection [J].
Chambers, Jeremy W. ;
Maguire, Tobi G. ;
Alwine, James C. .
JOURNAL OF VIROLOGY, 2010, 84 (04) :1867-1873