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KMID : 0578319950050060629
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Molecules and Cells
1995 Volume.5 No. 6 p.629 ~ p.633
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Effect of Human Cytomegalovirus immediate Early Gene Expression on Cytosolic Free Calcium concentration
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kang Jyung-hee
Lee Chan-Hee
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Abstract
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A sustained increase in cytosolic free calcium concentration ([Ca^(2+)]_(i)) was observed in human fibroblast cells infected with human cytomegalovirus (HCMV). The increase continued after the onset of HCMV major immediate early (MIE) genes, suggesting the possible involvement of MIE gene expression in the increase in [Ca^(2+)]_(i). To test this possibility, plasmid DNAs containing HCMV MIE genes were transfected into human embryonic lung (HEL) cells and the [Ca^(2+)]_(i) was measured indirectly by using fluorescent calcium indicator Fura-2. Transfection of HEL cells with plasmid DNA containing the entire region of MIE (MIE1+2) resulted in a substantial increase in [Ca^(2+)]_(i), while transfection with MIE1 or MIE2 gene alone had no significant effect on [Ca^(2+)]_(i). This increase in [Ca^(2+)]_(i) reached a maximum level at 40 h after transfection of MIE1+2 gene. To investigate if the [Ca^(2+)]_(i) increase in MIE gene-transfected cells was MIE gene-specific, cells were transfected with various concentrations of the MIE1+2 gene. The [Ca^(2+)]_(i) increased as MIE1+2 gene concentration increased, suggesting a direct effect of the MIE genes. The increase in [Ca^(2+)]_(i) following transfection of MIE gene was inhibited by treatment of cells with dantroline, an intracellular calcium blocker. But nifedipine, an extracellular calcium channel blocker, did not show any effect on [Ca^(2+)]_(i). It is suggested that intracellular calcium source is more important than extracellular calcium source for the [Ca^(2+)]_(i) increase in MIE gene-transfected cells.
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