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

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 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.

scholarly journals Generation and Characterization of Yeast Two-Hybrid cDNA Libraries Derived From Two Distinct Mouse Pluripotent Cell Types

2012 ◽  
Vol 54 (2) ◽  
pp. 228-237 ◽  
Author(s):  
Ying Zheng ◽  
Xiaoying Tan ◽  
Joanna Pyczek ◽  
Jessica Nolte ◽  
D. V. Krishna Pantakani ◽  
...  
Keyword(s):  
Cell Types ◽  
Cdna Libraries ◽  
Yeast Two Hybrid ◽  
Pluripotent Cell ◽  
Two Hybrid

scholarly journals The VPS33B-binding protein VPS16B is required in megakaryocyte and platelet α-granule biogenesis

Blood ◽  
2012 ◽  
Vol 120 (25) ◽  
pp. 5032-5040 ◽  
Author(s):  
Denisa Urban ◽  
Ling Li ◽  
Hilary Christensen ◽  
Fred G. Pluthero ◽  
Shao Zun Chen ◽  
...  
Keyword(s):  
Immunofluorescence Microscopy ◽  
Binding Protein ◽  
Yeast Two Hybrid ◽  
Binding Partner ◽  
Late Endosomes ◽  
Arc Syndrome ◽  
Membrane Bound ◽  
Golgi Network ◽  
Two Hybrid

Abstract Patients with platelet α or dense δ-granule defects have bleeding problems. Although several proteins are known to be required for δ-granule development, less is known about α-granule biogenesis. Our previous work showed that the BEACH protein NBEAL2 and the Sec1/Munc18 protein VPS33B are required for α-granule biogenesis. Using a yeast two-hybrid screen, mass spectrometry, coimmunoprecipitation, and bioinformatics studies, we identified VPS16B as a VPS33B-binding protein. Immunoblotting confirmed VPS16B expression in various human tissues and cells including megakaryocytes and platelets, and also in megakaryocytic Dami cells. Characterization of platelets from a patient with arthrogryposis, renal dysfunction, and cholestasis (ARC) syndrome containing mutations in C14orf133 encoding VPS16B revealed pale-appearing platelets in blood films and electron microscopy revealed a complete absence of α-granules, whereas δ-granules were observed. Soluble and membrane-bound α-granule proteins were reduced or undetectable, suggesting that both releasable and membrane-bound α-granule constituents were absent. Immunofluorescence microscopy of Dami cells stably expressing GFP-VPS16B revealed that similar to VPS33B, GFP-VPS16B colocalized with markers of the trans-Golgi network, late endosomes and α-granules. We conclude that VPS16B, similar to its binding partner VPS33B, is essential for megakaryocyte and platelet α-granule biogenesis.

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