Analysis of endoplasmic reticulum trafficking signals by combinatorial screening in mammalian cells. Proc Natl Acad Sci U S A 2001 Feb 27;98(5):2431-6
Date
02/28/2001Pubmed ID
11226256Pubmed Central ID
PMC30155DOI
10.1073/pnas.051630198Scopus ID
2-s2.0-0035956996 (requires institutional sign-in at Scopus site) 149 CitationsAbstract
To improve the accuracy of predicting membrane protein sorting signals, we developed a general methodology for defining trafficking signal consensus sequences in the environment of the living cell. Our approach uses retroviral gene transfer to create combinatorial expression libraries of trafficking signal variants in mammalian cells, flow cytometry to sort cells based on trafficking phenotype, and quantitative trafficking assays to measure the efficacy of individual signals. Using this strategy to analyze arginine- and lysine-based endoplasmic reticulum localization signals, we demonstrate that small changes in the local sequence context dramatically alter signal strength, generating a broad spectrum of trafficking phenotypes. Finally, using sequences from our screen, we found that the potency of di-lysine, but not di-arginine, mediated endoplasmic reticulum localization was correlated with the strength of interaction with alpha-COP.
Author List
Zerangue N, Malan MJ, Fried SR, Dazin PF, Jan YN, Jan LY, Schwappach BAuthor
Michael J. Malan MD Assistant Professor in the Anesthesiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceAnimals
Combinatorial Chemistry Techniques
Endoplasmic Reticulum
Flow Cytometry
Fluorescent Antibody Technique
Genes, Reporter
Golgi Apparatus
Molecular Sequence Data
Signal Transduction
Two-Hybrid System Techniques
