Effect of HIV type 1 Tat protein on butyric acid-induced differentiation in a hematopoietic progenitor cell line.
The trans-activator protein (Tat) of HIV-1 plays an important role in viral pathogenesis. Since Tat has been shown to alter expression of a number of host cellular genes, we have investigated the role of Tat in modulating gene expression and differentiation in hematopoietic progenitor cells. Tat protein was introduced in K562 cells, a human hematopoietic progenitor cell line, by either scrape-loading onto HeLa (HL)-tat cells or direct electroporation of an affinity-purified glutathione S-transferase (GST)-Tat fusion protein. Under these conditions, butyric acid-induced hemoglobin production in K562 cells was suppressed by 65 and 52%, respectively. However, coculturing with wild-type HeLa cells or electroporation with the control GST protein did not decrease hemoglobin production. To confirm the presence of bioactive Tat protein within K562 cells, the cells were transiently transfected with a pHIV/LTR-CAT prior to the introduction of Tat. A 30- to 40-fold induction in CAT gene expression was observed in the transfected K562 cells, which were either cocultured with HL-tat or were electroporated with GST-Tat. Simultaneous transient transfection of K562 cells with a TAR expression plasmid, to compete for the availability of Tat protein, significantly downregulated the HIV LTR trans-activation by Tat. In addition, overexpression of the TAR RNAs in K562 cells was able to downregulate the suppressive effect of Tat on butyric acid-induced differentiation. RT-PCR analysis of the total RNAs isolated from these cells demonstrated that Tat protein suppressed the butyric acid-induced gamma-globin gene expression by an average of 54% without affecting the level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNAs. These data indicate that the viral Tat protein plays a significant role in abrogating erythroid differentiation in K562 cells.
Mondal
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Department of Pharmacology, Tulane University School of Medicine, New Orleans, Louisiana 70112, USA.
Agrawal
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D002087:Butyrates / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D020148:Butyric Acid; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D015696:Gene Products, tat / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D005914:Globins / Q000096:biosynthesis; D016325:HIV Long Terminal Repeat / Q000187:drug effects; D015497:HIV-1 / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006454:Hemoglobins / Q000096:biosynthesis; D006801:Humans; D012313:RNA / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics / Q000494:pharmacology; D015533:Transcriptional Activation / Q000187:drug effects; D054322:tat Gene Products, Human Immunodeficiency Virus
D002087:Butyrates; D015696:Gene Products, tat; D006454:Hemoglobins; D015534:Trans-Activators; D054322:tat Gene Products, Human Immunodeficiency Virus; D020148:Butyric Acid; D012313:RNA; D005914:Globins