Facile manipulation of protein localization in fission yeast through GBP-GFP binding

Abstract

GFP-binding protein (or GBP) has been recently developed in various systems and organisms as an efficient tool to purify GFP fusion proteins. Due to the high affinity between GBP and GFP or GFP variants, this GBP-based approach is also ideally suited to alter the localization of functional proteins in live cells. To promote GBP-targeting approach to be more widely and easily used in fission yeast Schizosaccharomyces pombe, we developed a set of pFA6a-, pJK148-, and pUC119-based vectors containing GBP or GBP-mCherry coding sequences and inducible nmt or constitutive adh promoters with different strength. The GBP or GBP-mCherry fragments can serve as cassettes for N- or C-terminal genomic tagging of genes of interest. We illustrated the application of these vectors in construction of yeast strains with Dma1 or Cdc7 tagged with GBP-mCherry and efficient targeting of Dma1- or Cdc7-GBP-mCherry to SPB by Sid4-GFP. This series of vectors should help to facilitate the application of GBP-targeting approach in manipulating protein localization and analysis of gene function in fission yeast, on the level of single genes, as well as in systematic scale.