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Gene. 2016 Jul 1;585(1):65-70. doi: 10.1016/j.gene.2016.03.013. Epub 2016 Mar 19.

Investigating direct interaction between Escherichia coli topoisomerase I and RecA.

Author information

1
Department of Chemistry and Biochemistry, Florida International University, Miami, Florida, USA.
2
Department of Oncology, Georgetown University, Washington, District of Columbia, USA.
3
Department of Physics, Florida International University, Miami, Florida, USA.
4
Biomolecular Sciences Institute, Florida International University, Miami, Florida, USA.
#
Contributed equally

Abstract

Protein-protein interactions are of special importance in cellular processes, including replication, transcription, recombination, and repair. Escherichia coli topoisomerase I (EcTOP1) is primarily involved in the relaxation of negative DNA supercoiling. E. coli RecA, the key protein for homologous recombination and SOS DNA-damage response, has been shown to stimulate the relaxation activity of EcTOP1. The evidence for their direct protein-protein interaction has not been previously established. We report here the direct physical interaction between E. coli RecA and topoisomerase I. We demonstrated the RecA-topoisomerase I interaction via pull-down assays, and surface plasmon resonance measurements. Molecular docking supports the observation that the interaction involves the topoisomerase I N-terminal domains that form the active site. Our results from pull-down assays showed that ATP, although not required, enhances the RecA-EcTOP1 interaction. We propose that E. coli RecA physically interacts with topoisomerase I to modulate the chromosomal DNA supercoiling.

KEYWORDS:

DNA topoisomerase I; Molecular docking; Protein–protein interactions; Pull-down assay; RecA; SPR

PMID:
27001450
PMCID:
PMC4838544
DOI:
10.1016/j.gene.2016.03.013
[Indexed for MEDLINE]
Free PMC Article

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