Towards the Systematic Mapping and Engineering of the Protein Prenylation Machinery in Saccharomyces cerevisiae

Stein, Viktor, Kubala, Marta H., Steen, Jason, Grimmond, Sean M. and Alexandrov, Kirill (2015) Towards the Systematic Mapping and Engineering of the Protein Prenylation Machinery in Saccharomyces cerevisiae. PLoS One, 10 3: . doi:10.1371/journal.pone.0120716

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Author Stein, Viktor
Kubala, Marta H.
Steen, Jason
Grimmond, Sean M.
Alexandrov, Kirill
Title Towards the Systematic Mapping and Engineering of the Protein Prenylation Machinery in Saccharomyces cerevisiae
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2015-03
Year available 2015
Sub-type Article (original research)
DOI 10.1371/journal.pone.0120716
Open Access Status DOI
Volume 10
Issue 3
Total pages 15
Place of publication San Francisco, CA United States
Publisher Public Library of Science
Collection year 2016
Language eng
Formatted abstract
Protein prenylation is a widespread and highly conserved eukaryotic post-translational modification that endows proteins with the ability to reversibly attach to intracellular membranes. The dynamic interaction of prenylated proteins with intracellular membranes is essential for their signalling functions and is frequently deregulated in disease processes such as cancer. As a result, protein prenylation has been pharmacologically targeted by numerous drug discovery programs, albeit with limited success. To a large extent, this can be attributed to an insufficient understanding of the interplay of different protein prenyltransferases and the combinatorial diversity of the prenylatable sequence space. Here, we report a high-throughput, growth-based genetic selection assay in Saccharomyces cerevisiae based on the Ras Recruitment System which, for the first time, has allowed us to create a comprehensive map of prenylatable protein sequences in S. cerevisiae. We demonstrate that potential prenylatable space is sparsely (6.2%) occupied leaving room for creation of synthetic orthogonal prenylatable sequences. To experimentally demonstrate that, we used the developed platform to engineer mutant farnesyltransferases that efficiently prenylate substrate motives that are not recognised by endogenous protein prenyltransferases. These uncoupled mutants can now be used as starting points for the systematic engineering of the eukaryotic protein prenylation machinery.
Keyword Geranylgeranyltransferase Type I
Plasma Membrane Localization
A Factor
Substrate Recognition
Peptide Libraries
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
Institute for Molecular Bioscience - Publications
 
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