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Yeast

Title:		Facile synthesis of budding yeast a-factor and its use to synchronize cells of alpha mating type
Author(s):		O'Reilly N; Charbin A; Lopez-Serra L; Uhlmann F;
Address:		"Peptide Synthesis Laboratory, Cancer Research UK London Research Institute, UK"
Journal Title:		Yeast
Year:		2012
Volume:		20120529
Issue:		6
Page Number:		233 - 240
DOI:		10.1002/yea.2906
ISSN/ISBN:		1097-0061 (Electronic) 0749-503X (Linking)
Abstract:		"The ease with which populations of the budding yeast Saccharomyces cerevisiae can be synchronized using the mating pheromone alpha-factor has been invaluable for studies of the cell cycle. The alpha-factor response pathway has also remained an important model to study the molecular mechanism of G-protein coupled receptor signalling. alpha-Factor is a 13 amino acids long peptide that is readily available by automated peptide synthesis. However, only cells of the a mating type respond to alpha-factor. Cells of the opposite alpha mating type respond to a-factor, a farnesylated and C-terminally methylated 12 amino acids peptide. Because of its more difficult chemical synthesis, a-factor is not readily available and consequently the a-factor response is less well understood. Here we describe an improved strategy for producing a-factor, based on solid-phase peptide synthesis, followed by two simple steps in solution that show favourable characteristics and good yield. We demonstrate the successful use of the resulting a-factor to synchronize cell cycle progression of alpha cells. Notably, the a-factor concentrations required for cell synchronization are an order of magnitude lower than typically used alpha-factor concentrations. Despite a similar cell cycle response, shmoo formation was less pronounced compared to alpha-factor-treated a cells. Our protocol makes a-factor widely accessible, extending the ease of cell cycle synchronization to budding yeast cells of both mating types and facilitating the study of a-factor signalling"
Keywords:		Cell Cycle/*drug effects Mating Factor Mycology/*methods Peptides/*chemical synthesis/isolation & purification/*metabolism Saccharomyces cerevisiae/*growth & development/*metabolism;
Notes:		"MedlineO'Reilly, Nicola Charbin, Adrian Lopez-Serra, Lidia Uhlmann, Frank eng Research Support, Non-U.S. Gov't England 2012/05/30 Yeast. 2012 Jun; 29(6):233-40. doi: 10.1002/yea.2906. Epub 2012 May 29"