scholarly journals Subunit Oligomerization and Substrate Recognition of the Escherichia coli ClpYQ (HslUV) Protease Implicated by In Vivo Protein-Protein Interactions in the Yeast Two-Hybrid System

2004 ◽  
Vol 186 (7) ◽  
pp. 2224-2224
Author(s):  
Yi-Ying Lee ◽  
Chiung-Fang Chang ◽  
Chueh-Ling Kuo ◽  
Meng-Ching Chen ◽  
Chien Hung Yu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Hybrid System ◽  
Protein Interactions ◽  
Substrate Recognition ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

scholarly journals Subunit Oligomerization and Substrate Recognition of the Escherichia coli ClpYQ (HslUV) Protease Implicated by In Vivo Protein-Protein Interactions in the Yeast Two-Hybrid System

2003 ◽  
Vol 185 (8) ◽  
pp. 2393-2401 ◽  
Author(s):  
Yi-Ying Lee ◽  
Chiung-Fang Chang ◽  
Chueh-Ling Kuo ◽  
Meng-Ching Chen ◽  
Chien Hung Yu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Hybrid System ◽  
Protein Interactions ◽  
Large Subunit ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

ABSTRACT The Escherichia coli ClpYQ (HslUV) is an ATP-dependent protease that consists of an ATPase large subunit with homology to other Clp family ATPases and a peptidase small subunit related to the proteasomal β-subunits of eukaryotes. Six identical subunits of both ClpY and ClpQ self-assemble into an oligomeric ring, and two rings of each subunit, two ClpQ rings surrounded by single ClpY rings, form a dumbbell shape complex. The ClpYQ protease degrades the cell division inhibitor, SulA, and a positive regulator of capsule transcription, RcsA, as well as RpoH, a heat shock sigma transcription factor. Using the yeast-two hybrid system, we explored the in vivo protein-protein interactions of the individual subunits of the ClpYQ protease involved in self-oligomerization, as well as in recognition of specific substrates. Interactions were detected with ClpQ/ClpQ, ClpQ/ClpY, and ClpY/SulA. No interactions were observed in experiments with ClpY/ClpY, ClpQ/RcsA, and ClpQ/SulA. However, ClpY, lacking domain I (ClpYΔI) was able to interact with itself and with intact ClpY. The C-terminal region of ClpY is important for interaction with other ClpY subunits. The previously defined PDZ-like domains at the C terminus of ClpY, including both D1 and D2, were determined to be indispensable for substrate binding. Various deletion and random point mutants of SulA were also made to verify significant interactions with ClpY. Thus, we demonstrated in vivo hetero- and homointeractions of ClpQ and ClpY molecules, as well as a direct association between ClpY and substrate SulA, thereby supporting previous in vitro biochemical findings.

The Yeast Two-Hybrid System: A Tool for Mapping Protein–Protein Interactions

2015 ◽  
Vol 2015 (5) ◽  
pp. pdb.top083345 ◽  
Author(s):  
Jitender Mehla ◽  
J. Harry Caufield ◽  
Peter Uetz
Keyword(s):  
Hybrid System ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
System A ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

Determination of Protein-Protein Interactions of ICln by the Yeast Two-Hybrid System

2001 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Andreas Schmarda ◽  
Friedrich Fresser ◽  
Martin Gschwentner ◽  
Johannes Fürst ◽  
Markus Ritter ◽  
...  
Keyword(s):  
Hybrid System ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid
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