ABSTRACT
<p><b>OBJECTIVE</b>To study the role of a BTB/POZ domain protein in the expression of hsp90alpha gene.</p><p><b>METHODS</b>The eukaryotic expression plasmids of sense- and antisense-GAGA related protein (GRP) or empty vector were transfected into Jurkat cells with pREP4 episomal vector plasmids carrying the hsp90alpha promoter sequence from -1756 to +37 and control plasmids pMCAT. Total RNA was extracted. The relative promoter activity of hsp90alpha-CAT reporter gene was determined by competitive RT-PCR assay.</p><p><b>RESULTS</b>GRP markly increased the relative promoter activity of hsp90alpha-CAT reporter gene during heat shock.</p><p><b>CONCLUSION</b>GRP may promote the expression of hsp90alpha gene by participating in chromatin remolding.</p>
Subject(s)
Animals , Humans , Amino Acid Motifs , Checkpoint Kinase 1 , Cloning, Molecular , DNA , Metabolism , DNA, Complementary , Metabolism , DNA-Binding Proteins , Metabolism , Drosophila , Genetics , Drosophila Proteins , Genetics , Gene Expression Regulation , HSP90 Heat-Shock Proteins , Genetics , Heat-Shock Response , Genetics , Homeodomain Proteins , Chemistry , Genetics , Metabolism , Repressor Proteins , Genetics , Transcription Factors , Genetics , Two-Hybrid System TechniquesABSTRACT
<p><b>OBJECTIVE</b>To explore GAGA-like element binding protein in human cells.</p><p><b>METHODS</b>Yeast one-hybrid system was used to screen the GAGA-like element binding proteins in HTLV-1 transformed Jurkat cell cDNA fusion library. Total RNA extracted from Jurkat cells was first labeled by reverse transcription, and was taken as cDNA probe to hybridize with the candidate positive clones.</p><p><b>RESULTS</b>9 positive clones were obtained, and 6 out of the 9 clones were positively hybridized with the cDNA probe.</p><p><b>CONCLUSIONS</b>6 candidate clones encoding for GAGA-like element binding proteins were obtained from Jurkat cells for further investigation.</p>