TY - JOUR
T1 - YY1 and FoxD3 regulate antiretroviral zinc finger protein OTK18 promoter activation induced by HIV-1 infection
AU - Buescher, James L.
AU - Martinez, Lindsey B.
AU - Sato, Shinji
AU - Okuyama, Satoshi
AU - Ikezu, Tsuneya
N1 - Funding Information:
Acknowledgments We would like to thank NIH AIDS Research and Reference Reagent Program for pHIV-LTR-Luc and YU2 plasmids, Byran Cullen for pHIT/G plasmid, Edward Sato for human YY1 expression plasmid, and Robert Hromas for human FoxD3 (Genesis) expression plasmid, CNND Tissue and Cell Core Facility (Li Wu and Anuja Ghorpade) for primary culture of human microglia and monocyte elutriation, Michael Jacobsen and Meg Marquardt for editorial assistance, Santi Gorantla for consultation in viral pseudotyping, and Myhanh Che for viral titration. This work is funded by National Institute of Health R01 MH072539 (TI), P01 NS043985 (TI), and NCRR P20RR15635 (TI). The authors declare no conflict of interest or financial interests in this work.
PY - 2009/3
Y1 - 2009/3
N2 - OTK18 is a C2H2 type zinc finger protein involved in the regulation of HIV-1 replication in human mononuclear phagocytes. Previously, we reported OTK18 expression in brain perivascular macrophages but not in microglia in HIV encephalitis brain. We have cloned the OTK18 promoter region proximal to the transcriptional start site and determined the region responsible (-884/+1) for the basal transcriptional activity in a microglia cell line. Sequential deletion mutation analyses reveal three important response elements: Yingyang-1 (YY1; -805/-777), an HIV-1 response element for promoter activation; FoxD3 (-743/-725), a negative regulatory element; and Ets response element (-725/-707), a basal transcriptional activity response element. HIV-1 infection-induced upregulation of YY1 and c-Ets-1 protein, binding to the promoter region as determined by immunoblotting and chromatin immunoprecipitation and polymerase chain reaction (PCR) assays, and induction of YY1 was also observed in virus-infected monocyte-derived macrophages. Silencing of FoxD3 and YY1 in the cell line by small interfering RNA duplexes specific to these molecules significantly up- and downregulated basal OTK18 promoter activity in FoxD3 and YY1 response element-dependent manners, respectively. On the other hand, infection of primary cultured human microglia significantly reduced YY1 expression and induced FoxD3 as determined by immunoblotting and reverse transcription real-time PCR. These data suggest that HIV-1 induces OTK18 expression through a YY1-mediated manner in human macrophages, although its gene expression is suppressed by FoxD3 upregulation and YY1 downregulation in human microglia. This mechanism may explain the perivascular macrophage-specific expression of OTK18 in HIV encephalitis brains.
AB - OTK18 is a C2H2 type zinc finger protein involved in the regulation of HIV-1 replication in human mononuclear phagocytes. Previously, we reported OTK18 expression in brain perivascular macrophages but not in microglia in HIV encephalitis brain. We have cloned the OTK18 promoter region proximal to the transcriptional start site and determined the region responsible (-884/+1) for the basal transcriptional activity in a microglia cell line. Sequential deletion mutation analyses reveal three important response elements: Yingyang-1 (YY1; -805/-777), an HIV-1 response element for promoter activation; FoxD3 (-743/-725), a negative regulatory element; and Ets response element (-725/-707), a basal transcriptional activity response element. HIV-1 infection-induced upregulation of YY1 and c-Ets-1 protein, binding to the promoter region as determined by immunoblotting and chromatin immunoprecipitation and polymerase chain reaction (PCR) assays, and induction of YY1 was also observed in virus-infected monocyte-derived macrophages. Silencing of FoxD3 and YY1 in the cell line by small interfering RNA duplexes specific to these molecules significantly up- and downregulated basal OTK18 promoter activity in FoxD3 and YY1 response element-dependent manners, respectively. On the other hand, infection of primary cultured human microglia significantly reduced YY1 expression and induced FoxD3 as determined by immunoblotting and reverse transcription real-time PCR. These data suggest that HIV-1 induces OTK18 expression through a YY1-mediated manner in human macrophages, although its gene expression is suppressed by FoxD3 upregulation and YY1 downregulation in human microglia. This mechanism may explain the perivascular macrophage-specific expression of OTK18 in HIV encephalitis brains.
KW - HIV-1
KW - Macrophage
KW - Microglia
KW - Promoter regulation
KW - Transcription factor
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U2 - 10.1007/s11481-008-9139-x
DO - 10.1007/s11481-008-9139-x
M3 - Article
C2 - 19034670
AN - SCOPUS:60549107195
SN - 1557-1890
VL - 4
SP - 103
EP - 115
JO - Journal of Neuroimmune Pharmacology
JF - Journal of Neuroimmune Pharmacology
IS - 1
ER -