The Ras recruitment system, a novel approach to the study of protein–protein interactions

The Ras recruitment system, a novel approach to the study of protein–protein interactions The yeast two-hybrid system represents one of the most efficient approaches currently available for identifying and characterizing protein–protein interactions (1–4) . Although very powerful, this procedure exhibits several problems and inherent limitations (5) . A new system, the Sos recruitment system (SRS), was developed recently (6) based on a different readout from that of the two-hybrid system (6–8) . SRS overcomes several of the limitations of the two-hybrid system and thus serves as an attractive alternative for studying protein–protein interactions between known and novel proteins. Nevertheless, we encountered a number of problems using SRS and so have developed an improved protein recruitment system, designated the Ras recruitment system (RRS), based on the absolute requirement that Ras be localized to the plasma membrane for its function (9,10) . Ras membrane localization and activation can be achieved through interaction between two hybrid proteins. We have demonstrated the effectiveness of the novel RRS system using five different known protein–protein interactions and have identified two previously unknown protein–protein interactions through a library screening protocol. The first interaction (detailed here) is between JDP2, a member of the basic leucine zipper (bZIP) family, and C/EBP γ , a member of the CCAAT/enhancer-binding protein (C/EBP) family. The second interaction is between the p21-activated protein kinase Pak65 and a small G protein (described in the accompanying paper by Aronheim et al . (11) ). The RRS system significantly extends the usefulness of the previously described SRS system and overcomes several of its limitations. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Current Biology Elsevier

The Ras recruitment system, a novel approach to the study of protein–protein interactions

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Publisher
Elsevier
Copyright
Copyright © 1998 Elsevier Science Ltd
ISSN
0960-9822
D.O.I.
10.1016/S0960-9822(98)70467-1
Publisher site
See Article on Publisher Site

Abstract

The yeast two-hybrid system represents one of the most efficient approaches currently available for identifying and characterizing protein–protein interactions (1–4) . Although very powerful, this procedure exhibits several problems and inherent limitations (5) . A new system, the Sos recruitment system (SRS), was developed recently (6) based on a different readout from that of the two-hybrid system (6–8) . SRS overcomes several of the limitations of the two-hybrid system and thus serves as an attractive alternative for studying protein–protein interactions between known and novel proteins. Nevertheless, we encountered a number of problems using SRS and so have developed an improved protein recruitment system, designated the Ras recruitment system (RRS), based on the absolute requirement that Ras be localized to the plasma membrane for its function (9,10) . Ras membrane localization and activation can be achieved through interaction between two hybrid proteins. We have demonstrated the effectiveness of the novel RRS system using five different known protein–protein interactions and have identified two previously unknown protein–protein interactions through a library screening protocol. The first interaction (detailed here) is between JDP2, a member of the basic leucine zipper (bZIP) family, and C/EBP γ , a member of the CCAAT/enhancer-binding protein (C/EBP) family. The second interaction is between the p21-activated protein kinase Pak65 and a small G protein (described in the accompanying paper by Aronheim et al . (11) ). The RRS system significantly extends the usefulness of the previously described SRS system and overcomes several of its limitations.

Journal

Current BiologyElsevier

Published: Oct 8, 1998

References

  • Tag games in yeast: the two-hybrid system and beyond
    Boeke, J; Brachmann, RK
  • Comparison of thermosensitive alleles of the CDC25 gene involved in the cAMP metabolism of Saccharomyces cerevisiae
    Petitjean, A; Higler, F; Tatchell, K
  • Chp, a homologue of the GTPase Cdc42Hs, activates the JNK pathway and is implicated in reorganizing the actin cytoskeleton
    Aronheim, A; Broder, YC; Cohen, A; Fritsch, A; Belisle, B; Abo, A

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