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Molecular & Cellular Proteomics 6:413-424, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

The Alliance for Cellular Signaling Plasmid Collection

A Flexible Resource for Protein Localization Studies and Signaling Pathway Analysis^*,S

Joelle R. Zavzavadjian^,, Sam Couture^,, Wei Sun Park^,¶, James Whalen^,¶, Stephen Lyon^,||, Genie Lee^,, Eileen Fung^,, Qingli Mi^,, Jamie Liu^,, Estelle Wall^,, Leah Santat^,, Kavitha Dhandapani^,, Christine Kivork^,, Adrienne Driver^,, Xiaocui Zhu^,, Mi Sook Chang^,, Baljinder Randhawa^,, Elizabeth Gehrig^,¶, Heather Bryan^,¶, Mary Verghese^,¶, Andreia Maer^,||, Brian Saunders^,||, Yuhong Ning^,||, Shankar Subramaniam^,||, Tobias Meyer^,¶, Melvin I. Simon^,, Nancy O’Rourke^,¶, Grischa Chandy^,¶ and Iain D. C. Fraser^,,**

From the Alliance for Cellular Signaling, Molecular Biology Laboratory, California Institute of Technology, Pasadena, California 91125, ^¶ Microscopy Laboratory, Stanford University, Palo Alto, California 94305, and ^|| Bioinformatics and Data Coordination Laboratory, University of California San Diego, La Jolla, California 92093

Cellular responses to inputs that vary both temporally and spatially are determined by complex relationships between the components of cell signaling networks. Analysis of these relationships requires access to a wide range of experimental reagents and techniques, including the ability to express the protein components of the model cells in a variety of contexts. As part of the Alliance for Cellular Signaling, we developed a robust method for cloning large numbers of signaling ORFs into Gateway® entry vectors, and we created a wide range of compatible expression platforms for proteomics applications. To date, we have generated over 3000 plasmids that are available to the scientific community via the American Type Culture Collection. We have established a website at www.signaling-gateway.org/data/plasmid/ that allows users to browse, search, and blast Alliance for Cellular Signaling plasmids. The collection primarily contains murine signaling ORFs with an emphasis on kinases and G protein signaling genes. Here we describe the cloning, databasing, and application of this proteomics resource for large scale subcellular localization screens in mammalian cell lines.

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