Identifying proteins that affect mRNA localization in living cells.
Messenger RNA transport has emerged as a significant mechanism for regulating gene expression. Many of the protein factors affecting RNA transport remain unknown. The emergence of green fluorescent protein (GFP) fluorescence microscopy allows imaging in living cells and an increased understanding of in vivo molecular transport. GFP imaging is now applied to RNA transport by engineering RNA hairpins into the RNA of interest and observing fluorescence from GFP fused to an RNA-binding protein that recognizes the hairpins. In yeast, different genetic backgrounds can be tested to identify various proteins that affect RNA transport and localization. The technology also allows the swapping of different regions of the RNA to determine the cis requirements for transport. GFP RNA imaging opens many possibilities to examine RNA transport in real time in a variety of different organisms.
Brodsky
Alexander S
AS
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School and The Dana-Farber Cancer Institute, Boston, MA 02115, USA.
Silver
Pamela A
PA
D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism
C098733:ASH1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D012313:RNA