Bacterial Growth Inhibition Screen (BGIS): harnessing recombinant protein toxicity for rapid and unbiased interrogation of protein function

Haihong Guo, Nengwei Xu, Malte Prell, Hiltrud Königs, Benita Hermanns-Sachweh, Bernhard Lüscher, Ferdinand Kappes*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

In two proof-of-concept studies, we established and validated the Bacterial Growth Inhibition Screen (BGIS), which explores recombinant protein toxicity in Escherichia coli as a largely overlooked and alternative means for basic characterization of functional eukaryotic protein domains. By applying BGIS, we identified an unrecognized RNA-interacting domain in the DEK oncoprotein (this study) and successfully combined BGIS with random mutagenesis as a screening tool for loss-of-function mutants of the DNA modulating domain of DEK [1]. Collectively, our findings shed new light on the phenomenon of recombinant protein toxicity in E. coli. Given the easy and rapid implementation and wide applicability, BGIS will extend the repertoire of basic methods for the identification, analysis and unbiased manipulation of proteins.

Original languageEnglish
Pages (from-to)1422-1437
Number of pages16
JournalFEBS Letters
Volume595
Issue number10
DOIs
Publication statusPublished - May 2021

Keywords

  • DEK
  • Escherichia coli
  • RNA-binding domain
  • oncogene
  • protein domain
  • protein function
  • recombinant protein toxicity

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