Mechanism of presynaptic filament stabilization by the bacteriophage T4 UvsY recombination mediator protein

Biochemistry
Jie LiuScott W Morrical

Abstract

UvsY is the recombination mediator protein (RMP) of bacteriophage T4, which promotes homologous recombination by facilitating presynaptic filament assembly. The results of previous studies suggest that UvsY promotes the assembly of presynaptic filaments in part by stabilizing interactions between T4 UvsX recombinase and single-stranded DNA (ssDNA). To test this hypothesis, we studied the interactions of UvsX and UvsY with a fluorescein-derivatized oligonucleotide. This assay distinguishes between bipartite UvsX- or UvsY-ssDNA and tripartite UvsX-UvsY-ssDNA complex formation via differential fluorescence quenching effects. Salt stabilities of the three complexes were measured at equilibrium in the presence and absence of various nucleotide ligands of the UvsX protein and also under steady-state conditions for UvsX-catalyzed ssDNA-dependent ATP hydrolysis. The results demonstrate that UvsY globally stabilizes UvsX-ssDNA complexes, consistent with an increase in the apparent equilibrium binding affinity, K(ss)omega, of the UvsX-ssDNA interactions. The UvsY-mediated affinity increase is observed at equilibrium in the presence of ADP, ATPgammaS, or in the absence of the nucleotide and also at steady-state in the presence of ATP. Int...Continue Reading

References

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Citations

Apr 30, 2013·DNA Repair·William J GrahamCynthia A Haseltine
Dec 17, 2008·Proceedings of the National Academy of Sciences of the United States of America·Eli RothenbergTaekjip Ha
Feb 27, 2009·Nucleic Acids Research·Joshua N Farb, Scott W Morrical
Oct 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Hang XuScott W Morrical
Dec 7, 2010·Virology Journal·Jie Liu, Scott W Morrical
Jun 21, 2008·Journal of Molecular Biology·Kiran PantMark C Williams
Apr 8, 2000·Trends in Biochemical Sciences·K N Kreuzer
May 24, 2011·Critical Reviews in Biochemistry and Molecular Biology·Jie LiuScott W Morrical

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