4.5 Article

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

Journal

FEBS LETTERS
Volume 595, Issue 10, Pages 1422-1437

Publisher

WILEY
DOI: 10.1002/1873-3468.14072

Keywords

DEK; Escherichia coli; oncogene; protein domain; protein function; recombinant protein toxicity; RNA‐ binding domain

Funding

  1. China Scholarship Council
  2. Deutsche Forschungsgemeinschaft [DFG] [KA 2799/1, LU 466/16-1/2]
  3. START Program of the Faculty of Medicine, RWTH Aachen University

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The BGIS method investigates recombinant protein toxicity in E. coli, identifying protein domains and enabling screening. The study discovered an unrecognized RNA-interacting domain in the DEK oncoprotein, and successfully used BGIS for screening loss-of-function mutants in the DNA modulating domain.
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.

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