Karyoplasmic interaction selection strategy: A general strategy to detect protein-protein interactions in mammalian cells

E. R. Fearon, T. Finkel, M. L. Gillison, S. P. Kennedy, J. F. Casella, G. F. Tomaselli, J. S. Morrow, C. Van Dang

Research output: Contribution to journalArticle

69 Scopus citations

Abstract

We describe a strategy and reagents for study of protein-protein interactions in mammalian cells, termed the karyoplasmic interaction selection strategy (KISS). With this strategy, specific protein-protein interactions are identified by reconstitution of the functional activity of the yeast transcriptional activator GAL4 and the resultant transcription of a GAL4-regulated reporter gene. Reconstitution of GAL4 function results from specific interaction between two chimeric proteins: one contains the DNA- binding domain of GAL4; the other contains a transcriptional activation domain. Transcription of the reporter gene occurs if the two chimeric proteins can form a complex that reconstitutes the DNA-binding and transcriptional activation functions of GAL4. Using the KISS system, we demonstrate specific interactions for sequences from three different pairs of proteins that complex in the cytoplasm. In addition, we demonstrate that reporter genes encoding cell surface or drug-resistance markers can be specifically activated as a result of protein-protein interactions. With these selectable markers, the KISS system can be used to screen specialized cDNA libraries to identify novel protein interactions.

Original languageEnglish (US)
Pages (from-to)7958-7962
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number17
DOIs
StatePublished - Jan 1 1992
Externally publishedYes

Keywords

  • Chinese hamster ovary cells
  • GAL4 protein
  • herpes simplex virus protein VP16
  • hygromycin resistance

ASJC Scopus subject areas

  • General

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