DOI: 10.1073/pnas.96.17.9533. Studying interactions of four proteins in the yeast two-hybrid system: Structural resemblance of the pVHL/elongin BC/hCUL-2 complex with the ubiquitin ligase complex SKP1/cullin/F-box protein

Studying interactions of four proteins in the yeast two-hybrid system: Structural resemblance of the pVHL/elongin BC/hCUL-2 complex with the ubiquitin ligase complex SKP1/cullin/F-box protein

10.1073/pnas.96.17.9533

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The yeast two-hybrid system is a powerful technique that detects interactions between two proteins and has been useful in identifying new binding partners. However, the system fails to detect protein–protein interactions that require the presence of additional components of a multisubunit complex. Here we demonstrate that the vector YIpDCE1 can be used to express elongins B and C in yeast, and that these proteins form a stable complex that interacts with the von Hippel–Lindau tumor-suppressor gene product (pVHL). Only when pVHL and elongins B and C (VBC) are present does an interaction with the cullin family member, hCUL-2, occur, forming the heterotetrameric pVHL/elongin BC/hCUL-2 complex. This system was then used to map the binding region of hCUL-2 for the VBC complex. The first amino-terminal 108 aa of hCUL-2 are necessary for interaction with the VBC complex. The elongin BC dimer acts as a bridge between pVHL and hCUL-2 because pVHL and hCUL-2 can form distinct complexes with elongins B and C. These results reveal a striking structural resemblance of pVHL/elongin BC/hCUL-2 complex with the E3-like ubiquitin ligase complex SKP1/Cullin/F-box protein with respect to protein composition and sites of interactions. Thus, it seems possible that pVHL/elongin BC/hCUL-2 complex will possess ubiquitin ligase activity targeting specific proteins for degradation by the proteasome.

Bibliography

Pause, A., Peterson, B., Schaffar, G., Stearman, R., & Klausner, R. D. (1999). Studying interactions of four proteins in the yeast two-hybrid system: Structural resemblance of the pVHL/elongin BC/hCUL-2 complex with the ubiquitin ligase complex SKP1/cullin/F-box protein. Proceedings of the National Academy of Sciences, 96(17), 9533â9538.

Authors 5

Arnim Pause (first)
Brett Peterson (additional)
Gregor Schaffar (additional)
Robert Stearman (additional)
Richard D. Klausner (additional)

References 34 Referenced 73

10.1097/00005792-199711000-00001
J R Gnarra, D R Duan, Y Weng, J S Humphrey, D Y Chen, S Lee, A Pause, C F Dudley, F Latif, I Kuzmin, et al. Biochim Biophys Acta 1242, 201–210 (1996). / Biochim Biophys Acta by Gnarra J R (1996)
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10.1126/science.7660130
T Kishida, T M Stackhouse, F Chen, M I Lerman, B Zbar Cancer Res 55, 4544–4548 (1995). / Cancer Res by Kishida T (1995)
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J W Conaway, T Kamura, R C Conaway Biochim Biophys Acta 1377, M49–54 (1998). / Biochim Biophys Acta by Conaway J W (1998)
10.1016/S0092-8674(00)81267-2
10.1073/pnas.95.3.993
G Siemeister, K Weindel, K Mohrs, B Barleon, G Martiny-Baron, D Marme Cancer Res 56, 2299–2301 (1996). / Cancer Res by Siemeister G (1996)
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J J Michel, Y Xiong Cell Growth Differ 9, 435–449 (1998). / Cell Growth Differ by Michel J J (1998)
10.1016/S0378-1119(98)00179-6
10.1074/jbc.272.43.27444
M A Osborne, S Dalton, J P Kochan Bio/Technology 13, 1474–1478 (1995). / Bio/Technology by Osborne M A (1995)
10.1016/S0958-1669(97)80029-8
10.1074/jbc.272.37.22995
10.1126/science.279.5354.1216
10.1126/science.279.5354.1219
10.1073/pnas.95.14.7987

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:41 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 4:47 p.m.)
Indexed	3 weeks, 5 days ago (Aug. 5, 2025, 8:42 a.m.)
Issued	26 years ago (Aug. 17, 1999)
Published	26 years ago (Aug. 17, 1999)
Published Online	26 years ago (Aug. 17, 1999)
Published Print	26 years ago (Aug. 17, 1999)

Funders 0

None

BibTeX

@article{Pause_1999, title={Studying interactions of four proteins in the yeast two-hybrid system: Structural resemblance of the pVHL/elongin BC/hCUL-2 complex with the ubiquitin ligase complex SKP1/cullin/F-box protein}, volume={96}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.96.17.9533}, DOI={10.1073/pnas.96.17.9533}, number={17}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Pause, Arnim and Peterson, Brett and Schaffar, Gregor and Stearman, Robert and Klausner, Richard D.}, year={1999}, month=aug, pages={9533–9538} }

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