Cloning and characterization of hGMEB1, a novel glucocorticoid modulatory element binding protein

• Published in: FEBS letters Vol. 452; no. 3; pp. 170 - 176
• Main Authors: Thériault, Jimmy R, Charette, Steve J, Lambert, Herman, Landry, Jacques
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 11.06.1999
• Subjects: 3T3 Cells; Amino Acid Sequence; Animals; Binding Sites; Cell Line; Cloning, Molecular; Cricetinae; Glucocorticoid modulatory element binding protein; Glutathione Transferase - biosynthesis; Heat-Shock Proteins; HeLa Cells; HSP27; HSP27 Heat-Shock Proteins; Humans; Mice; Molecular Sequence Data; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Oligodeoxyribonucleotides - chemistry; Oligodeoxyribonucleotides - metabolism; Protein Biosynthesis; Rats; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - metabolism; Saccharomyces cerevisiae; Sequence Alignment; Sequence Homology, Amino Acid; Transcription factor; Transcription Factors - chemistry; Transcription Factors - genetics; Transcription Factors - metabolism; Transfection; Transcription factor; Glucocorticoid modulatory element binding protein; HSP27
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(99)00634-1

A 21-bp element called glucocorticoid modulatory element (GME) modulates the glucocorticoid receptor-mediated responses via the binding of an as yet poorly characterized trans-acting complex of proteins containing the 88-kDa GMEB1 and the 67-kDa GMEB2. Using heat shock protein 27 (HSP27) as bait in the yeast two-hybrid assay, we cloned a 1.83-kb cDNA encoding a novel 573-amino acid protein called human GMEB1 (hGMEB1). hGMEB1 possesses a KDWK domain, contains sequences almost identical (36/38) to three tryptic peptides of rat GMEB1 and shares 38% identity with rat GMEB2. hGMEB1 is ubiquitously expressed as a 85-kDa protein in all cell lines and tissues examined. In vitro translated hGMEB1 bound specifically to GME oligonucleotides yielding a complex of similar size to the complex obtained using rat liver nuclear extracts. Both complexes were supershifted with an antibody specific to hGMEB1. Co-immunoprecipitation experiments confirmed the in vivo interaction of HSP27 with hGMEB1.