|
| Status |
Public on Aug 26, 2015 |
| Title |
HT1080 WT or STAT deficient exposed to IFN beta or human trophoblast medium |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
Previous microarray data indicated a significant increase in interferon (IFN) stimulated gene expression in cells exposed to conditioned medium from primary human trophoblasts compared to non conditioned medium. Here we want to compare the effects of conditioned media, non conditioned media, and non conditioned media spiked with 100 units of IFN beta in wild type HT1080 cells as well as in HT1080 cells deficient in STAT1 (a key signaling molecular in the IFN pathway).
Interferon stimulated genes (ISGs) target viruses at various stages of their infectious life cycles, including at the earliest stage of viral entry. Here we identify ArfGAP with dual pleckstrin homology (PH) domains 2 (ADAP2) as a gene upregulated by type I IFN treatment in a STAT1-dependent manner. ADAP2 functions as a GTPase-activating protein (GAP) for Arf6 and binds to phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) and PI(3,4)P2. We show that overexpression of ADAP2 suppresses dengue virus (DENV) and vesicular stomatitis virus (VSV) infection in an Arf6 GAP activity-dependent manner, while exerting no effect on coxsackievirus B (CVB) or Sendai virus (SeV) replication. We further show that ADAP2 expression induces macropinocytosis and that ADAP2 strongly associates with actin-enriched membrane ruffles and with Rab8a- and LAMP1-, but not EEA1- or Rab7-, positive vesicles. Utilizing two techniques—light-sensitive neutral red (NR)-containing DENV and fluorescence assays for virus internalization—we show that ADAP2 primarily restricts DENV infection at the stage of virion entry and/or intracellular trafficking and that incoming DENV and VSV particles associate with ADAP2 during their entry. Taken together, this study identifies ADAP2 as an ISG that exerts antiviral effects against RNA viruses by altering Arf6-mediated trafficking to disrupt viral entry.
|
| |
|
| Overall design |
compare the cells exposed to CM and the cells exposed to NC media spiked with IFN to the control NC exposed cells, in wild type (2f-TGH) vs Stat -/- cells (HT1080U3A)
|
| |
|
| Contributor(s) |
l Sadovsky Y, Coyne C |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Aug 25, 2015 |
| Last update date |
Oct 11, 2016 |
| Contact name |
Yoel Sadovsky |
| E-mail(s) |
ysadovsky@mwri.magee.edu
|
| Phone |
412-641-2675
|
| Organization name |
MAGEE WOMENS RESEARCH INSTITUTE
|
| Department |
Obstetrics and Gynecology
|
| Lab |
Sadovsky
|
| Street address |
204 Craft Ave
|
| City |
PITTSBURGH |
| State/province |
Pennsylvania |
| ZIP/Postal code |
15213 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL14550 |
Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Probe Name Version) |
|
| Samples (8)
|
|
| Relations |
| BioProject |
PRJNA293806 |
Less...
More...