The ras recruitment system, a novel approach to the study of protein-protein interactions - PubMed (original) (raw)
The ras recruitment system, a novel approach to the study of protein-protein interactions
Y C Broder et al. Curr Biol. 1998.
Free article
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/EBPgamma, 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.
Similar articles
- The Ras Recruitment System (RRS) for the Identification and Characterization of Protein-Protein Interactions.
Aronheim A. Aronheim A. Methods Mol Biol. 2018;1794:61-73. doi: 10.1007/978-1-4939-7871-7_5. Methods Mol Biol. 2018. PMID: 29855951 - 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. Aronheim A, et al. Curr Biol. 1998 Oct 8;8(20):1125-8. doi: 10.1016/s0960-9822(98)70468-3. Curr Biol. 1998. PMID: 9778532 - cDNA library screening using the SOS recruitment system.
Huang W, Wang SL, Lozano G, de Crombrugghe B. Huang W, et al. Biotechniques. 2001 Jan;30(1):94-8, 100. doi: 10.2144/01301st06. Biotechniques. 2001. PMID: 11196326 - A family of C/EBP-related proteins capable of forming covalently linked leucine zipper dimers in vitro.
Williams SC, Cantwell CA, Johnson PF. Williams SC, et al. Genes Dev. 1991 Sep;5(9):1553-67. doi: 10.1101/gad.5.9.1553. Genes Dev. 1991. PMID: 1884998 - Yeast genetic methods for the detection of membrane protein interactions: potential use in drug discovery.
Fetchko M, Auerbach D, Stagljar I. Fetchko M, et al. BioDrugs. 2003;17(6):413-24. doi: 10.2165/00063030-200317060-00004. BioDrugs. 2003. PMID: 14614764 Review.
Cited by
- Specific Disruption of Ras2 CAAX Proteolysis Alters Its Localization and Function.
Ravishankar R, Hildebrandt ER, Greenway G, Asad N, Gore S, Dore TM, Schmidt WK. Ravishankar R, et al. Microbiol Spectr. 2023 Feb 14;11(1):e0269222. doi: 10.1128/spectrum.02692-22. Epub 2023 Jan 5. Microbiol Spectr. 2023. PMID: 36602340 Free PMC article. - Yeast Two-Hybrid System for Mapping Novel Dengue Protein Interactions.
Conde JN. Conde JN. Methods Mol Biol. 2022;2409:119-132. doi: 10.1007/978-1-0716-1879-0_9. Methods Mol Biol. 2022. PMID: 34709639 - The Methods Employed in Mass Spectrometric Analysis of Posttranslational Modifications (PTMs) and Protein-Protein Interactions (PPIs).
Yakubu RR, Nieves E, Weiss LM. Yakubu RR, et al. Adv Exp Med Biol. 2019;1140:169-198. doi: 10.1007/978-3-030-15950-4_10. Adv Exp Med Biol. 2019. PMID: 31347048 Free PMC article. Review. - Saccharomyces cerevisiae as a Tool to Investigate Plant Potassium and Sodium Transporters.
Locascio A, Andrés-Colás N, Mulet JM, Yenush L. Locascio A, et al. Int J Mol Sci. 2019 Apr 30;20(9):2133. doi: 10.3390/ijms20092133. Int J Mol Sci. 2019. PMID: 31052176 Free PMC article. Review. - WDR62 mediates TNFα-dependent JNK activation via TRAF2-MLK3 axis.
Prinz E, Aviram S, Aronheim A. Prinz E, et al. Mol Biol Cell. 2018 Oct 1;29(20):2470-2480. doi: 10.1091/mbc.E17-08-0504. Epub 2018 Aug 9. Mol Biol Cell. 2018. PMID: 30091641 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases