Specific protein-DNA and protein-protein interaction in the hig gene system, a plasmid-borne proteic killer gene system of plasmid Rts1

Plasmid
Q B TianT Hayashi

Abstract

The hig (host inhibition of growth) gene system of plasmid Rts1 belongs to the plasmid-encoded proteic killer gene family. Among the proteic killer genes described so far, hig is unique in that the toxin gene (higB) exists upstream of the antidote gene (higA). There are two promoters in the hig locus, Phig and PhigA, and only the former, which expresses both higB and higA genes, is negatively controlled by HigA and HigB proteins. In this study, we purified HigA protein by means of GST fusion. The electrophoretic mobility shift assay using the purified protein revealed that HigA specifically bound to the Phig region, but not to PhigA. The HigA-binding sequence was determined by DNase I footprinting assay to be a 56-bp sequence that completely covered the -35 and -10 boxes of Phig. The presence of two inverted repeats in the binding sequence and the identification of a dimer form of HigA by cross-linking experiment suggested that the protein bound to the Phig region as a dimer. HigB was purified as a GST fusion protein as well, though it was achieved only in the presence of HigA. HigA and GST-HigB formed a highly stable complex where the two proteins were present in an equimolar ratio.

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Citations

May 3, 2005·Nature Reviews. Microbiology·Kenn GerdesAnders Løbner-Olesen
Jun 30, 2010·Journal of Bacteriology·Amanda S Fivian-Hughes, Elaine O Davis
Jul 19, 2011·Journal of Bacteriology·Wai Ting ChanChew Chieng Yeo
May 25, 2002·Journal of Bacteriology·Takahiro MurataTetsuya Hayashi
Mar 4, 2005·Journal of Bacteriology·Xueyan Zhao, Roy David Magnuson
Oct 6, 2006·Molecular Microbiology·Mikkel Christensen-Dalsgaard, Kenn Gerdes
Nov 20, 2013·Mobile Genetic Elements·Simon J UnterholznerWilfried Rozhon
Sep 8, 2016·Biochemical and Biophysical Research Communications·Jingsi YangQuansheng Liu
Feb 23, 2019·Molecular Microbiology·Marc A SchureckChristine M Dunham
Nov 22, 2013·The Journal of Biological Chemistry·Marc A SchureckChristine M Dunham
Jul 6, 2016·Nucleic Acids Research·Marc A SchureckChristine M Dunham
Sep 10, 2019·Structure·Melek Cemre ManavDitlev Egeskov Brodersen

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