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Abstract
Cold shock domain (CSD) family members have been shown to play roles in either transcriptional activation or repression of many genes in various cell types. We have previously shown that CSD proteins dbpAv and dbpB (also known as YB-1) act to repress granulocyte-macrophage colony-stimulating factor transcription in human embryonic lung (HEL) fibroblasts via binding to single-stranded DNA regions across the promoter. Here we show that the same CSD factors are involved in granulocyte-macrophage colony-stimulating factor transcriptional activation in Jurkat T cells. Unlike the mechanisms of CSD repression in HEL fibroblasts, CSD-mediated activation in Jurkat T cells is not mediated through DNA binding but presumably through protein-protein interactions via the C terminus of the CSD protein with transcription factors such as RelA/NF-kappaB p65. We demonstrate that Jurkat T cells lack truncated CSD factor subtypes present in HEL fibroblasts, which raises the possibility that the cellular content of CSD proteins may determine their final role as activators or repressors of transcription.
Highlights
Cold shock domain (CSD)1 proteins were originally identified in bacteria and have been shown to be highly conserved throughout evolution from bacteria to humans [1,2,3]
Detection of CSD Complexes in Jurkat T Cells—We have previously shown that both human embryonic lung (HEL) fibroblasts and HUT78 T cells contain nuclear CSD proteins and that they bind to the human granulocyte-macrophage colony-stimulating factor (GM-CSF) proximal promoter [21,22,23]
To determine whether Jurkat T cells contained CSD proteins, Jurkat T cell nuclear extract was bound in a gel shift assay to single-stranded oligonucleotides spanning domain 1 and domain 2 of the human proximal GM-CSF promoter (Fig. 1B) and compared with HUT78 T cell CSD factor binding on the same oligonucleotides
Summary
Cold shock domain (CSD)1 proteins were originally identified in bacteria and have been shown to be highly conserved throughout evolution from bacteria to humans [1,2,3]. Detection of CSD Complexes in Jurkat T Cells—We have previously shown that both HEL fibroblasts and HUT78 T cells contain nuclear CSD proteins and that they bind to the human GM-CSF proximal promoter [21,22,23].
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