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Science 31 July 1992:
Vol. 257. no. 5070, pp. 680 - 682
DOI: 10.1126/science.1496382
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Articles

Science, Vol 257, Issue 5070, 680-682
Copyright © 1992 by American Association for the Advancement of Science

articles

A protein kinase substrate identified by the two-hybrid system

X Yang, EJ Hubbard, and M Carlson

Department of Genetics and Development, College of Physicians and Surgeons, Columbia University, New York, NY 10032.

A genetic method, the two-hybrid system, was used to identify four genes encoding proteins that interact with the SNF1 protein kinase from yeast. One of the genes, SIP1, was independently isolated as a multicopy suppressor of defects caused by reduced SNF1 kinase activity, and genetic evidence supports its function in the SNF1 pathway. The SIP1 protein co-immunoprecipitated with SNF1 and was phosphorylated in vitro. Thus, the two-hybrid system, which is applicable to any cloned gene, can be used to detect physical interactions between protein kinases and functionally related substrate proteins.


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