Protein-protein interactions directing resolvase site-specific recombination: a structure-function analysis.

Hughes, Robert E.; Rice, Phoebe A.; Steitz, Thomas A.; Grindley, Nigel D. F.

doi:10.1002/j.1460-2075.1993.tb05788.x

Cited by 47 publications

(53 citation statements)

References 39 publications

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“…2, our intense efforts to detect recombination on standard substrates for inversion were unsuccessful. With inference from the crystal structure of gd resolvase, residues within region B most likely belong to an a helix that constitutes the solution dimer interface and, in addition, to a so-called 'linker' region that connects the DNA-binding and the catalytic domain (Hughes et al 1990(Hughes et al , 1993Sanderson et al 1990;Grindley 1994;Yang & Steitz 1995). Remarkably, 63% of the residues within region B are shared between ISXc5 resolvase and C-C Liu et al…”

Section: Discussionmentioning

confidence: 99%

The resolvase encoded by Xanthomonas campestris transposable element ISXc5 constitutes a new subfamily closely related to DNA invertases

Liu

Hühne

et al. 1998

Genes to Cells

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Background: Conservative site-specific recombination is responsible for the resolution of cointegrates which result during the transposition of class II transposable elements. Resolution is catalysed by a transposonencoded recombinase, resolvase, that belongs to a large family of recombinases, including DNA invertases. Resolvases and the related invertases are likely to employ similar reaction mechanisms during recombination. There are important differences, however. Resolvases require two accessory DNA binding sites within each of the two directly repeated recombination sites. Invertases instead need a host factor, Fis, and an enhancer type DNA sequence, in addition to two inversely orientated recombination sites.

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Section: Discussionmentioning

confidence: 99%

The resolvase encoded by Xanthomonas campestris transposable element ISXc5 constitutes a new subfamily closely related to DNA invertases

Liu

Hühne

et al. 1998

Genes to Cells

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“…After introduction of the Fis-independent mutation M114V, the inversion activity of this mutant is exactly the same as that of the M114V mutant itself. Residue M-108 is located in the ␣-helix, which in resolvase is involved in the dimerization of the protein (6,29). This ␣-helix might therefore be the target for Fis interaction.…”

Section: Vol 177 1995 Gin Mutant Suppression By a Fis-independent Mmentioning

confidence: 99%

Gin mutants that can be suppressed by a Fis-independent mutation

et al. 1995

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The Gin invertase of bacteriophage Mu mediates recombination between two inverted gix sites. Recombination requires the presence of a second protein, Fis, which binds to an enhancer sequence. We have isolated 24 different mutants of Gin that are impaired in DNA inversion but proficient in DNA binding. Six of these mutants could be suppressed for inversion by introduction of a second mutation, which when present in the wild-type gin gene causes a Fis-independent phenotype. Only one of the six resulting double mutants shows an inversion efficiency which is comparable to that of the wild-type Gin and which is independent of Fis. The corresponding mutation, M to I at position 108 (M108I), is located in a putative ␣-helical structure, which in the homologous ␥␦ resolvase has been implicated in dimerization. The properties of the M108I mutant suggest that in Gin this dimerization helix might also be the target for Fis interaction. The five other mutants that show a restored inversion after introduction of a Fis-independent mutation appear to be completely dependent on Fis for this inversion. The corresponding mutations are located in different domains of the protein. The properties of these mutants in connection with the role of Fis in inversion will be discussed.

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“…The I1 10R and V114R mutants of y6 resolvase were overexpressed in the AR120 E. coli strain from pRH57 and pNG264 (Hughes et al, 1993). The yS resolvase and mutant proteins were purified as described previously (Hatfull et al, 1989).…”

Section: Plasmidsmentioning

confidence: 99%

Secondary and tertiary structural changes in γδ resolvase: Comparison of the wild‐type enzyme, the I11OR mutant, and the C‐terminal DNA binding domain in solution

et al. 1997

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y6Resolvase is a site-specific DNA recombinase (MI 20.5 kDa) in Escherichia coli that shares homology with a family of bacterial resolvases and invertases. We have characterized the secondary and tertiary structural behavior of the cloned DNA binding domain (DBD) and a dimerization defective mutant in solution. Low-salt conditions were found to destabilize the tertiary structure of the DBD dramatically, with concomitant changes in the secondary structure that were localized near the hinge regions between the helices. The molten tertiary fold appears to contribute significantly to productive DNA interactions and supports a mechanism of DNA-induced folding of the tertiary structure, a process that enables the DBD to adapt in conformation for each of the three imperfect palindromic sites. At high salt concentrations, the monomeric I1 1OR resolvase shows a minimal perturbation to the three helices of the DBD structure and changes in the linker segment in comparison to the cloned DBD containing the linker. Comparative analysis of the NMR spectra suggest that the I1 1OR mutant contains a folded catalytic core of -60 residues and that the segment from residues 100 to 149 are devoid of regular structure in the IllOR resolvase. No increase in the helicity of the linker region of IllOR resolvase occurs on binding DNA. These results support a subunit rotation model of strand exchange that involves the partial unfolding of the catalytic domains.Keywords: DNA; folding; interaction; NMR; protein; recombination; resolvase; structure y6 Resolvase (MI 20.5 kDa) is a site-specific DNA recombinase in Escherichia coli that shares homology with a family of resolvases and invertases in a variety of bacteria (Grindley & Reed, 1985). The structure of resolvase can be divided into three distinct regions, consisting of a catalytic domain (residues 1-120), a DNA binding domain (residues 149-183), and a linker region between the two globular domains. Limited proteolysis of 715 resolvase yields a C-terminal 43-residue fragment (5 kDa) with DNA binding activity and an N-terminal 140-residue fragment (15.5 kDa) with oligomerization and catalytic functions. The structure of the catalytic domain has been determined by X-ray crystallography

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Protein-protein interactions directing resolvase site-specific recombination: a structure-function analysis.

Cited by 47 publications

References 39 publications

The resolvase encoded by Xanthomonas campestris transposable element ISXc5 constitutes a new subfamily closely related to DNA invertases

The resolvase encoded by Xanthomonas campestris transposable element ISXc5 constitutes a new subfamily closely related to DNA invertases

Gin mutants that can be suppressed by a Fis-independent mutation

Secondary and tertiary structural changes in γδ resolvase: Comparison of the wild‐type enzyme, the I11OR mutant, and the C‐terminal DNA binding domain in solution

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