4.5 Article

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

期刊

FEBS LETTERS
卷 595, 期 10, 页码 1422-1437

出版社

WILEY
DOI: 10.1002/1873-3468.14072

关键词

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

资金

  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

向作者/读者索取更多资源

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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