A constraint network of interactions: protein–protein interaction analysis of the yeast type II phosphatase Ptc1p and its adaptor protein Nbp2p

Abstract

The Slt2p-Slt2p interaction reported in Fig. 8E was mistakenly not included in Table 1. The authors apologise for this error.

A fluorescent reporter for mapping cellular protein‐protein interactions in time and space

“…Nap1p is localized in the cytosol, the nucleus, and the bud neck. We recently confirmed N ub ‐Kcc4p as binding partner of Nap1CRU (Hruby et al , 2011). Kcc4p is a septin‐localized protein kinase (Barral et al , 1999; Okuzaki and Nojima, 2001).…”

“…To test the workflow for transferring interactions revealed by large‐scale interaction experiments into single‐cell analysis, we screened Net1CRU against an array of 382 different N ub fusions (N ub array) (Hruby et al , 2011). Among others Ubc9p, Cdc14p, and Fkh1p were identified as further Net1p‐binding partners (Table I; Visintin et al , 1999).…”

“…Large scale Split‐Ub assays were performed as described (Hruby et al , 2011; Dünkler et al , 2012). Measuring interactions between individual N ub ‐ and C ub ‐fusion proteins by spotting yeast cells expressing both fusions onto 5‐FOA and SD Ura‐containing media was essentially as described (Eckert and Johnsson, 2003).…”

“…N ub ‐ and C ub ‐fusion genes were constructed as described (Hruby et al , 2011; Dünkler et al , 2012). Gene deletions were performed by PCR‐based methods as described (Janke et al , 2004).…”

“…A mechanistic understanding of cellular biology requires a comprehensive knowledge about the protein interactions of the cell (Uetz et al , 2000; Gavin et al , 2002; Krogan et al , 2006; Yu et al , 2008; Breitkreutz et al , 2010). Split‐protein sensors comprise a family of related techniques that contributed in small‐ and large‐scale experiments to this cumulative endeavour (Miller et al , 2005; Tarassov et al , 2008; Hruby et al , 2011; Lowder et al , 2011; Stynen et al , 2012). The Split‐Ubiquitin method (Split‐Ub) is the prototype of these techniques (Müller and Johnsson, 2008; Stynen et al , 2012).…”