Identification and Characterization of Interacting Partners of Rab GTPases by Yeast Two-Hybrid Analyses

2008 ◽  
pp. 111-125 ◽  
Author(s):  
Mark Kail ◽  
Angelika Barnekow
Keyword(s):  
Rab Gtpases ◽  
Yeast Two Hybrid ◽  
Identification And Characterization ◽  
Two Hybrid

Identification and Characterization of Inhibitors That Specifically Target TORC2 in Multiple Myeloma

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 5026-5026
Author(s):  
Bao Hoang ◽  
Joseph Gera ◽  
Alan Lichtenstein
Keyword(s):  
Multiple Myeloma ◽  
Conflicts Of Interest ◽  
Yeast Two Hybrid ◽  
Drug Induced ◽  
Akt Phosphorylation ◽  
Protein Substrates ◽  
Yeast Two Hybrid System ◽  
Identification And Characterization ◽  
Two Hybrid

Abstract Abstract 5026 Recent data indicate that TORC2 is necessary for survival of multiple myeloma (MM) cells. Currently, drugs targeting mTOR either inhibit TORC1 alone or both TORC1 and TORC2. To identify drugs that specifically target TORC2, we performed a drug screen in a yeast-two hybrid system to identify compounds that prevented an interaction between rictor and mTOR. We identified several potential compounds and have begun to characterize their molecular activity. These compounds induced significant MM cell apoptosis when used at concentrations below 4 uM. This cytotoxic effect was accomplished by a down regulation of TOR activity which was specific for TORC2 (ie., decreased S473 AKT phosphorylation and NDRG T346 phosphorylation). Co-immunoprecipitation experiments confirmed that at least some of the compounds prevented binding of rictor to mTOR within MM cells while having no effect on binding of raptor to mTOR. In addition, myeloma cells expressing TORC2 phosphomimetic protein substrates (AKTS473D or SGKS422D) were significantly less sensitive to apoptosis as compared to the empty vector control when treated with these compounds. These data suggest that the drug-induced cytotoxicity was mediated specifically through the inhibition of TORC2 kinase activity. These results are the initial characterization of TORC2-specific drugs and support a rationale for targeting TORC2 in multiple myeloma. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Identification and characterization of two novel SH2 domain-containing proteins from a yeast two hybrid screen with the ABL tyrosine kinase

Oncogene ◽  
1997 ◽  
Vol 15 (11) ◽  
pp. 1255-1262 ◽  
Author(s):  
Tsukasa Oda ◽  
Jody Kujovich ◽  
Margaret Reis ◽  
Brenda Newman ◽  
Brian J Druker
Keyword(s):  
Tyrosine Kinase ◽  
Sh2 Domain ◽  
Yeast Two Hybrid ◽  
Abl Tyrosine Kinase ◽  
Identification And Characterization ◽  
Two Hybrid

Selection and characterization of large collections of peptide aptamers through optimized yeast two-hybrid procedures

2006 ◽  
Vol 1 (3) ◽  
pp. 1066-1091 ◽  
Author(s):  
Marc B T Bickle ◽  
Eric Dusserre ◽  
Olivier Moncorgé ◽  
Hélène Bottin ◽  
Pierre Colas
Keyword(s):  
Yeast Two Hybrid ◽  
Hybrid Procedures ◽  
Peptide Aptamers ◽  
Two Hybrid

scholarly journals A novel yeast two-hybrid approach to identify CDPK substrates: Characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein1

FEBS Letters ◽  
2006 ◽  
Vol 580 (3) ◽  
pp. 904-911 ◽  
Author(s):  
Miguel A. Rodriguez Milla ◽  
Yuichi Uno ◽  
Ing-Feng Chang ◽  
Jared Townsend ◽  
Eileen A. Maher ◽  
...  
Keyword(s):  
Zinc Finger ◽  
Hybrid Approach ◽  
Yeast Two Hybrid ◽  
Two Hybrid

scholarly journals Construction and characterization of a normalized yeast two-hybrid library derived from a human protein-coding clone collection

BioTechniques ◽  
2008 ◽  
Vol 44 (2) ◽  
pp. 265-273 ◽  
Author(s):  
Jorja Degrado-Warren ◽  
Max Dufford ◽  
Jian Chen ◽  
Paul L. Bartel ◽  
Donna Shattuck ◽  
...  
Keyword(s):  
Human Protein ◽  
Yeast Two Hybrid ◽  
Protein Coding ◽  
Two Hybrid

scholarly journals Schizosaccharomyces pombeRdh54 (TID1) Acts with Rhp54 (RAD54) to Repair Meiotic Double-Strand Breaks

2003 ◽  
Vol 14 (11) ◽  
pp. 4707-4720 ◽  
Author(s):  
Michael G. Catlett ◽  
Susan L. Forsburg
Keyword(s):  
Meiotic Recombination ◽  
Double Strand Breaks ◽  
Cell Analysis ◽  
Yeast Two Hybrid ◽  
Spore Viability ◽  
Strand Breaks ◽  
Functional Homology ◽  
Damage Repair ◽  
Two Hybrid

We report the characterization of rdh54+, the second fission yeast Schizosaccharomyces pombe Rad54 homolog. rdh54+shares sequence and functional homology to budding yeast RDH54/TID1. Rdh54p is present during meiosis with appropriate timing for a meiotic recombination factor. It interacts with Rhp51 and the meiotic Rhp51 homolog Dmc1 in yeast two-hybrid assays. Deletion of rdh54+has no effect on DNA damage repair during the haploid vegetative cell cycle. In meiosis, however, rdh54Δ shows decreased spore viability and homologous recombination with a concomitant increase in sister chromatid exchange. The rdh54Δ single mutant repairs meiotic breaks with similar timing to wild type, suggesting redundancy of meiotic recombination factors. Consistent with this, the rdh54Δ rhp54Δ double mutant fails to repair meiotic double strand breaks. Live cell analysis shows that rdh54Δ rhp54Δ asci do not arrest, but undergo both meiotic divisions with near normal timing, suggesting that failure to repair double strand breaks in S. pombe meiosis does not result in checkpoint arrest.

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