Bacterial Growth Inhibition Screen (BGIS) identifies a loss-of-function mutant of the DEK oncogene, indicating DNA modulating activities of DEK in chromatin

FEBS Letters

article

Haihong Guo¹  Malte Prell¹  Hiltrud Königs²  Nengwei Xu³  Tanja Waldmann⁴  Benita Hermans-Sachweh²  Elisa Ferrando-May⁵  Bernhard Lüscher¹  Ferdinand Kappes¹ 

[1] Institute for Biochemistry and Molecular Biology, Medical School, RWTH Aachen University;Institute of Pathology, Medical School, RWTH Aachen University;Department of Biological Sciences, Xi’an Jiaotong-Liverpool University;Doerenkamp-Zbinden Chair for In Vitro Toxicology and Biomedicine, University of Konstanz;Bioimaging Center, Department of Biology, University of Konstanz

关键词: chromatin;    SAP domain;    DEK;    Escherichia coli;    oncogene;    protein domain;    protein function;    recombinant protein toxicity;   

DOI  :  10.1002/1873-3468.14070

来源: John Wiley & Sons Ltd.

【 摘 要 】

The DEK oncoprotein regulates cellular chromatin function via a number of protein–protein interactions. However, the biological relevance of its unique pseudo-SAP/SAP-box domain, which transmits DNA modulating activities in vitro, remains largely speculative. As hypothesis-driven mutations failed to yield DNA-binding null (DBN) mutants, we combined random mutagenesis with the Bacterial Growth Inhibition Screen (BGIS) to overcome this bottleneck. Re-expression of a DEK-DBN mutant in newly established human DEK knockout cells failed to reduce the increase in nuclear size as compared to wild type, indicating roles for DEK–DNA interactions in cellular chromatin organization. Our results extend the functional roles of DEK in metazoan chromatin and highlight the predictive ability of recombinant protein toxicity in E. coli for unbiased studies of eukaryotic DNA modulating protein domains.

【 授权许可】

Unknown