Multi-probe RPA template sets to study RNA modulation and transcriptional control of BH3-only members of the Bcl-2 family.
The Bcl-2 family of apoptotic regulators comprises both pro- and anti-apoptotic proteins. Pro-apoptotic BH3-only members of the Bcl-2 family are subject to stringent control in normal proliferating cells, but can be activated by a variety of mechanisms in response to diverse apoptotic signals. To facilitate the simultaneous identification of BH3-only targets that are regulated at the RNA level, we have designed and assembled multi-probe ribonuclease protection assay templates and demonstrated their ability to detect multiple BH3-only transcripts in a variety of human cell lines and primary tissues. Following this, we have determined the modulation of BH3-only transcripts to a variety of apoptotic stimuli, in selected primary and transformed cell lines. Early experiments, showing that a given cell type upregulates a unique subset of BH3-only RNAs in response to different death-inducing stimuli, indicate that the multi-probe BH3-only RPA template sets could serve as useful diagnostic and analytical tools in cancer research.
Wang
Dehua
D
Johns Hopkins University, Department of Pediatric Oncology, Baltimore, MD, USA.
McDonnell
Maureen A
MA
Kelekar
Ameeta
A
D017209:Apoptosis; D051023:BH3 Interacting Domain Death Agonist Protein; D001681:Biological Assay / Q000379:methods; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D020869:Gene Expression Profiling; D019254:Genes, bcl-2 / Q000502:physiology*; D006801:Humans; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D012313:RNA / Q000096:biosynthesis* / Q000502:physiology*; D012260:Ribonucleases / Q000378:metabolism; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D002352:Carrier Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012313:RNA; D012260:Ribonucleases