« Previous Abstract"Identification of signature volatiles to discriminate Candida albicans, glabrata, krusei and tropicalis using gas chromatography and mass spectrometry"    Next AbstractEarly childhood lower respiratory illness and air pollution »

Yeast

Title:		In vivo selectively infective phage as a tool to detect protein interactions: evaluation of a novel vector system with yeast Ste7p-Fus3p interacting proteins
Author(s):		Hertveldt K; Robben J; Volckaert G;
Address:		"Laboratorium voor Gentechnologie, Kasteelpark Arenberg 21, B-3001 Leuven, Belgium. kirsten.Hertveldt@agr.kuleuven.ac.be"
Journal Title:		Yeast
Year:		2002
Volume:		19
Issue:		6
Page Number:		499 - 508
DOI:		10.1002/yea.857
ISSN/ISBN:		0749-503X (Print) 0749-503X (Linking)
Abstract:		"The selectively infective phage (SIP) approach allows rapid identification of interacting proteins by linking protein-protein interaction to phage infectivity. Infection of E. coli by filamentous phage depends on viral g3p. This protein consists of three domains, N1, N2 and CT. Phages lacking the N1 domain are non-infective unless a bait (X)-prey (Y) interaction links it to phage anchored N2-CT domains. We have developed all the vectors required for an in vivo selectively infective phage strategy (SIP). This includes a bait vector, pG3N1, a prey vector, pHOS41, and a gene III deletion helper phage, HPd3. The bait vector pG3N1 allows expression of a bait protein (X) fused to the C-terminus of the N1 domain. The prey vector pHOS41 allows expression of prey (Y) proteins, fused to the N-terminus of the N2-CT domains. The gene III deletion helper phage delivers all phage proteins necessary for phage production, except g3p. Escherichia coli transformed with these three vectors produces non-infective phages unless a bait-prey interaction links the g3p domains. Fus3p and Ste7p, two proteins from the Saccharomyces cerevisiae pheromone-responsive pathway have been cloned to evaluate the SIP strategy. The presence of the interacting N1-Fus3p adapter increased the infectivity of Ste7p-N2-CT phages approximately 1400-fold, which makes SIP a promising technology for the detection and further investigation of interacting proteins"
Keywords:		Capsid Proteins *Coliphages *Cytoskeletal Proteins DNA-Binding Proteins/genetics Escherichia coli/genetics/*virology Fungal Proteins/metabolism *Genetic Vectors Mitogen-Activated Protein Kinase Kinases Mitogen-Activated Protein Kinases/*metabolism Protein;
Notes:		"MedlineHertveldt, Kirsten Robben, Johan Volckaert, Guido eng Research Support, Non-U.S. Gov't England 2002/03/29 Yeast. 2002 Apr; 19(6):499-508. doi: 10.1002/yea.857"