Free article by Nobel Laureate in November MCP.’The monthly journal, Molecular and Cellular Proteomics (MCP), has made an article published by Nobel Laureate Dr. Andrew Z. Fire available free to the public on its website. The article, which appears in the November issue of the journal, is entitled “A Differential Cytolocalization Assay for Analysis of Macromolecular Assemblies in the Eukaryotic Cytoplasm.”
Dr. Fire was awarded the 2006 Nobel Prize in Physiology or Medicine along with Dr. Craig C. Mello for their discovery of RNA interference (RNAi). In the MCP paper, Fire and colleagues report on their development of an assay that allows them to observe the interactions between proteins in vivo.
The researchers tested the assay in C. elegans, using it to probe interactions among proteins involved in RNA interference (RNAi) and nonsense mediated decay (NMD) pathways. Several previously documented interactions were confirmed with the assay, and many new ones were observed. Fire et al. also used the assay to test a subset of the RNAi and NMD interactions in animals mutant for proteins central to each mechanism, and were able to identify several key associations that occurred in the improperly functioning processes.’
A Differential Cytolocalization Assay for Analysis of Macromolecular Assemblies in the Eukaryotic Cytoplasm. Abstract: We have developed a differential cytolocalization assay (DCLA) that allows the observation of cytoplasmic protein:protein interactions in vivo. In the DCLA assay, interactions are visualized as a relocalization of a GFP-tagged “prey” by a membrane bound “bait”. This assay has been tested and utilized in C. elegans to probe interactions among proteins involved in RNA interference (RNAi) and nonsense mediated decay (NMD) pathways. Several previously documented interactions have been confirmed with DCLA, while uniformly negative results were obtained in several controls in which no interaction was expected. Novel interactions were also observed including the association of SMG-5, a protein required for NMD, to several components of the RNAi pathway. The DCLA assay can be readily carried out under diverse conditions, allowing a dynamic assessment of protein interactions in vivo. We have used this property to test a subset of the RNAi and NMD interactions in animals mutant for proteins central to each mechanism, identifying several key associations in each machinery that can occur in vivo in the absence of a functional process. (Download PDF)
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Founded in 1906, the Society is based in Bethesda, Maryland, on the campus of the Federation of American Societies for Experimental Biology. The Society’s purpose is to advance the science of biochemistry and molecular biology through publication of scientific and educational journals (Journal of Biological Chemistry, Molecular and Cellular Proteomics, Journal of Lipid Research, Biochemistry and Molecular Biology Education), organization of scientific meetings, advocacy for funding of basic research and education, support of science education at all levels, and promoting the diversity of individuals entering the scientific workforce.’