期刊
METHODS
卷 58, 期 4, 页码 392-399出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2012.07.015
关键词
Y2H; Ribonucleotide reductase; DNA polymerase; MntR/PerR complex; Phage lambda; Proteasome
资金
- NIH [R01GM079710]
- NSF [1048199]
- Direct For Computer & Info Scie & Enginr
- Division of Computing and Communication Foundations [1048199] Funding Source: National Science Foundation
Protein complexes are typically analyzed by affinity purification and subsequent mass spectrometric analysis. However, in most cases the structure and topology of the complexes remains elusive from such studies. Here we investigate how the yeast two-hybrid system can be used to analyze direct interactions among proteins in a complex. First we tested all pairwise interactions among the seven proteins of Escherichia coli DNA polymerase III as well as an uncharacterized complex that includes MntR and PerR. Four and seven interactions were identified in these two complexes, respectively. In addition, we review Y2H data for three other complexes of known structure which serve as gold-standards, namely Varicella Zoster Virus (VZV) ribonucleotide reductase (RNR), the yeast proteasome, and bacteriophage lambda. Finally, we review an Y2H analysis of the human spliceosome which may serve as an example for a dynamic mega-complex. (c) 2012 Elsevier Inc. All rights reserved.
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