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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Jun 22, 2013 |
| Title |
Identification of Adaptive mutations in the influenza A virus non-structural 1 gene that increase cytoplasmic localization and differentially regulate host gene expression |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
The NS1 protein of influenza A virus (IAV) is a multifunctional virulence factor. Mouse adaptive mutations in the NS1 protein of the human isolate A/Hong Kong/1/1968(H3N2) (HK) have been previously reported to increase virulence, viral fitness, and interferon antagonism, but differ in binding to post-transcriptional processing factor CPSF30. Because nuclear trafficking is a major genetic determinant of influenza virus host adaptation, we assessed subcellular localization and host gene expression of NS1 adaptive mutations. Recombinant HK viruses with adaptive mutations in the NS1 gene were assessed for NS1 protein subcellular localization in mouse and human cells using confocal microscopy and cellular fractionation. HK-wt virus NS1 partitioned equivalently between the cytoplasm and nucleus in human cells but was defective in cytoplasmic localization in mouse cells. The adaptive mutations either increased the proportion or abundance of NS1 in the cytoplasm, and/or the nucleus. NS1 mutations that increased cytoplasmic distribution identified a putative second nuclear export signal (NES) spanning aa positions 98-106 LSEDWFMLM, (mutation sites in bold); with the strongest effect seen for mutation M106I. The putative NES in the NS3 protein was associated with cytoplasmic localization. The host gene expression profile of the adaptive mutants was determined by microarray analysis of infected mouse cells to show either high or low gene regulation (HGR or LGR) phenotypes that mapped to the amino-terminal and the carboxy-terminal regions respectively. The HGR and LGR mutations were predominantly down regulating versus up regulating respectively. The greatest effect on host gene expression in the HGR group correlated with the ability of the NS1 protein to bind CPSF30. To our knowledge this is the first report of roles of adaptive NS1 mutations that affect intracellular localization and regulation of host gene expression.
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| Overall design |
biological triplicates of mock infected mouse M1 cells; cells infected with A/HK/1/1968(H3N2) wt and NS1 mtuatn with mutaions, D2N, V23A, L98S, L98S + M106I, F103L, M106V, M106V + M124I, D125G, V180A, V226I, M106I, and R227K.
Cells were infected at a multiplicty of infection of 2 and cels were incubated for 8 hr at 37 C for 8 hrs before RNA extraction and analysis relative to mock PBS infected cells.
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| Contributor(s) |
Brown EG, Pelchat M, Forbes N |
| Citation(s) |
24391972 |
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| Submission date |
Jun 21, 2013 |
| Last update date |
Mar 04, 2019 |
| Contact name |
Earl Garnet Brown |
| E-mail(s) |
[email protected]
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| Phone |
613 224 7533
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| Organization name |
University of Ottawa
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| Department |
Biochemistry Microbiology and Immunology
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| Lab |
RGN 4109
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| Street address |
451 Smyth Rd
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| City |
Ottawa |
| State/province |
Ontario |
| ZIP/Postal code |
KIH 8M5 |
| Country |
Canada |
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| Platforms (1) |
| GPL6246 |
[MoGene-1_0-st] Affymetrix Mouse Gene 1.0 ST Array [transcript (gene) version] |
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| Samples (42)
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| GSM1172656 |
M1, HK-NS1 WT 1 |
| GSM1172657 |
M1, HK-NS1 WT 2 |
| GSM1172658 |
M1, HK-NS1 WT 3 |
| GSM1172659 |
M1, NS1 F103L 1 |
| GSM1172660 |
M1, NS1 F103L 2 |
| GSM1172661 |
M1, NS1 F103L 3 |
| GSM1172662 |
M1, NS1 M106V 1 |
| GSM1172663 |
M1, NS1 M106V 2 |
| GSM1172664 |
M1, NS1 M106V 3 |
| GSM1172665 |
M1, NS1 M106I 1 |
| GSM1172666 |
M1, NS1 M106I 2 |
| GSM1172667 |
M1, NS1 M106I 3 |
| GSM1173007 |
M1, NS1 D2N 1 |
| GSM1173008 |
M1, NS1 D2N 2 |
| GSM1173009 |
M1, NS1 D2N 3 |
| GSM1173010 |
M1, NS1 V23A 1 |
| GSM1173011 |
M1, NS1 V23A 2 |
| GSM1173012 |
M1, NS1 V23A 3 |
| GSM1173013 |
M1, NS1 L98S 1 |
| GSM1173014 |
M1, NS1 L98S 2 |
| GSM1173015 |
M1, NS1 L98S 3 |
| GSM1173016 |
M1, NS1 L98S/M106I 1 |
| GSM1173017 |
M1, NS1 L98S/M106I 2 |
| GSM1173018 |
M1, NS1 L98S/M106I 3 |
| GSM1173019 |
M1, NS1 M106V/M124I 1 |
| GSM1173020 |
M1, NS1 M106V/M124I 2 |
| GSM1173021 |
M1, NS1 M106V/M124I 3 |
| GSM1173022 |
M1, NS1 D125G 1 |
| GSM1173023 |
M1, NS1 D125G 2 |
| GSM1173024 |
M1, NS1 D125G 3 |
| GSM1173025 |
M1, NS1 V180A 1 |
| GSM1173026 |
M1, NS1 V180A 2 |
| GSM1173027 |
M1, NS1 V180A 3 |
| GSM1173028 |
M1, NS1 V226I 1 |
| GSM1173029 |
M1, NS1 V226I 2 |
| GSM1173030 |
M1, NS1 V226I 3 |
| GSM1173031 |
M1, NS1 R227K 1 |
| GSM1173032 |
M1, NS1 R227K 2 |
| GSM1173033 |
M1, NS1 R227K 3 |
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| Relations |
| BioProject |
PRJNA209217 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE48217_RAW.tar |
179.5 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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