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

FEBS Letters, vol. 595, pp. 1422–1437

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.

Authors 7

1. Haihong Guo Aachen Medical School Institute for Biochemistry and Molecular Biology Medical School Institute for Biochemistry and Molecular Biology

   RWTH Aachen University

   Affiliation as printed

   Institute for Biochemistry and Molecular Biology Medical School RWTH Aachen University Germany

   Institute for Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Germany

2. Nengwei Xu

   Xi’an Jiaotong-Liverpool University

   Affiliation as printed

   Department of Biological Sciences Suzhou Dushu Lake Science and Education Innovation District Suzhou Industrial Park Xi’an Jiaotong‐Liverpool University Suzhou China

   Department of Biological Sciences, Suzhou Dushu Lake Science and Education Innovation District, Suzhou Industrial Park, Xi'an Jiaotong-Liverpool University, Suzhou, China

3. Malte Prell Aachen Medical School Institute for Biochemistry and Molecular Biology Medical School Institute for Biochemistry and Molecular Biology

   RWTH Aachen University

   Affiliation as printed

   Institute for Biochemistry and Molecular Biology Medical School RWTH Aachen University Germany

   Institute for Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Germany

4. Hiltrud Königs Aachen Institute of Pathology Medical School

   RWTH Aachen University

   Affiliation as printed

   Institute of Pathology Medical School RWTH Aachen University Germany

   Institute of Pathology, Medical School, RWTH Aachen University, Germany

5. Benita Hermanns‐Sachweh Aachen Institute of Pathology Medical School

   RWTH Aachen University

   Affiliation as printed

   Institute of Pathology Medical School RWTH Aachen University Germany

   Institute of Pathology, Medical School, RWTH Aachen University, Germany

6. Bernhard Lüscher Aachen Medical School Institute for Biochemistry and Molecular Biology Medical School Institute for Biochemistry and Molecular Biology

   RWTH Aachen University

   Affiliation as printed

   Institute for Biochemistry and Molecular Biology Medical School RWTH Aachen University Germany

   Institute for Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Germany

7. Ferdinand Kappes corresponding Aachen Medical School Institute for Biochemistry and Molecular Biology Medical School Institute for Biochemistry and Molecular Biology

   RWTH Aachen University · Xi’an Jiaotong-Liverpool University

   Affiliation as printed

   Department of Biological Sciences Suzhou Dushu Lake Science and Education Innovation District Suzhou Industrial Park Xi’an Jiaotong‐Liverpool University Suzhou China

   Institute for Biochemistry and Molecular Biology Medical School RWTH Aachen University Germany

   Department of Biological Sciences, Suzhou Dushu Lake Science and Education Innovation District, Suzhou Industrial Park, Xi'an Jiaotong-Liverpool University, Suzhou, China

   Institute for Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Germany