ABSTRACT
Acquisition of heat shock factor 2 (HSF2) DNA binding activity is accompanied by induced transcription of heat shock genes in hemin-treated K562 cells undergoing erythroid differentiation. Previous studies revealed that HSF2 consists of two alternatively spliced isoforms, HSF2-alpha and HSF2-beta, whose relative abundance is developmentally regulated and varies between different tissues. To investigate whether the molar ratio of HSF2-alpha and HSF2-beta isoforms is crucial for the activation of HSF2 and whether the HSF2 isoforms play functionally distinct roles during the hemin-mediated erythroid differentiation, we generated cell clones expressing different levels of HSF2-alpha and HSF2-beta. We show that in parental K562 cells, the HSF2-alpha isoform is predominantly expressed and HSF2 can be activated upon hemin treatment. In contrast, when HSF2-beta is expressed at levels exceeding those of endogenous HSF2-alpha, the hemin-induced DNA binding activity and transcription of heat shock genes are repressed, whereas overexpression of HSF2-alpha results in an enhanced hemin response. Furthermore, the hemin-induced accumulation of globin, known as a marker of erythroid differentiation, is decreased in cells overexpressing HSF2-beta. We suggest that HSF2-beta acts as a negative regulator of HSF2 activity during hemin-mediated erythroid differentiation of K562 cells.
Subject(s)
Cell Differentiation/physiology , Erythrocytes/cytology , Gene Expression Regulation/physiology , Heat-Shock Proteins/genetics , Hemin/physiology , Transcription Factors/genetics , Alternative Splicing , Humans , Tumor Cells, CulturedABSTRACT
Partial inhibition of DNA synthesis stimulates the production of inorganic diphosphatase in Escherichia coli but the changes in diphosphate (PPi) level observed did not correlate with the enzyme activity. An accumulation of PPi was observed in the presence of inhibitors of RNA synthesis or nucleotide synthesis. In the former case the level of the enzyme did not change but in the latter case it increased. Thus the amount of inorganic diphosphatase alone does not determine the concentration of PPi in E. coli.
Subject(s)
Diphosphates/metabolism , Escherichia coli/enzymology , Pyrophosphatases/metabolism , DNA Replication/drug effects , Escherichia coli/drug effects , Escherichia coli/genetics , Fluorouracil/pharmacology , Inorganic Pyrophosphatase , Kinetics , Mutation , Nalidixic Acid/pharmacology , Pyrophosphatases/genetics , Transcription, Genetic/drug effects , Uracil/analogs & derivatives , Uracil/pharmacologyABSTRACT
Pyrophosphate (PPi) content of Escherichia coli is increased manyfold when the growth of the cells is limited by inhibition of the synthesis of nucleotides. The accumulated PPi is immediately degraded when inhibition is released and growth allowed to resume. Other tested nutritional deficiencies (starvation of carbon source, sulfate or an amino acid) fail to induce PPi accumulation.
