Quantitative proteomic analyses of influenza virus-infected cultured human lung cells.
Because they are obligate intracellular parasites, all viruses are exclusively and intimately dependent upon host cells for replication. Viruses, in turn, induce profound changes within cells, including apoptosis, morphological changes, and activation of signaling pathways. Many of these alterations have been analyzed by gene arrays, which measure the cellular "transcriptome." Until recently, it has not been possible to extend comparable types of studies to globally examine all the host cellular proteins, which are the actual effector molecules. We have used stable isotope labeling by amino acids in cell culture (SILAC), combined with high-throughput two-dimensional (2-D) high-performance liquid chromatography (HPLC)/mass spectrometry, to determine quantitative differences in host proteins after infection of human lung A549 cells with human influenza virus A/PR/8/34 (H1N1) for 24 h. Of the 4,689 identified and measured cytosolic protein pairs, 127 were significantly upregulated at >95% confidence, 153 were significantly downregulated at >95% confidence, and a total of 87 proteins were upregulated or downregulated more than 5-fold at >99% confidence. Gene ontology and pathway analyses indicated differentially regulated proteins and included those involved in host cell immunity and antigen presentation, cell adhesion, metabolism, protein function, signal transduction, and transcription pathways.
Coombs
Kevin M
KM
Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba R3E 0J6, Canada. kcoombs@cc.umanitoba.ca
Berard
Alicia
A
Xu
Wanhong
W
Krokhin
Oleg
O
Meng
Xiaobo
X
Cortens
John P
JP
Kobasa
Darwyn
D
Wilkins
John
J
Brown
Earl G
EG
D000596:Amino Acids / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015536:Down-Regulation; D054884:Host-Pathogen Interactions / Q000502:physiology*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype; D007251:Influenza, Human / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D007554:Isotopes; D008168:Lung / Q000378:metabolism / Q000473:pathology / Q000821:virology; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D015854:Up-Regulation; D014764:Viral Proteins / Q000378:metabolism
D000596:Amino Acids; D007554:Isotopes; D020543:Proteome; D014764:Viral Proteins