Studies of the dimerization and domain structure of gamma delta resolvase

Liu, T.; Liu, D.; DeRose, Eugene F.; Mullen, Gregory P.

doi:10.1016/s0021-9258(19)85422-4

Cited by 13 publications

(4 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our observation that Gin behaves as a dimer in solution during the gel filtration experiment is in agreement with the results found earlier with gel filtration experiments under comparable conditions for the homologous Hin invertase (Glasgow et al, 1989) and the rather homologous yd resolvase (Liu et al, 1993). For Hin almost the same value of Vo/Ve is found as in our experiments with Gin.…”

Section: Discussionsupporting

confidence: 93%

Gin Invertase of Bacteriophage .mu. is a Dimer in Solution, with the Domain for Dimerization in the N-Terminal Part of the Protein

Spaeny-Dekking

Hemert²,

P³

et al. 1995

Biochemistry

View full text Add to dashboard Cite

The Gin protein of bacteriophage Mu mediates recombination between two inverted repeat sequences. Gin binds as a dimer to each of these recombination sites. We show that Gin is a dimer in solution also, and that the dimerization is probably stabilized by hydrophobic interactions between the subunits. The subunits of the dimer could efficiently be cross-linked with the 4-A cross-linker diepoxybutane. Spontaneous oxidation of Cys(24) and/or Cys(27) also resulted in intersubunit cross-linking. One or both cysteine residues are located at the interface of the Gin dimer, which maps the dimerization domain in the N-terminal part of the protein. Binding of the disulfide-bonded dimers of Gin to a recombination site was strongly reduced, suggesting that the subunits need to reorient in order to form a stable protein-DNA complex. In the protein-DNA complex, however, oxidation of cysteine residues still seems to be possible, indicating that the N-terminal parts of two Gin subunits are also in close proximity when bound to DNA.

show abstract

Section: Discussionsupporting

confidence: 93%

Gin Invertase of Bacteriophage .mu. is a Dimer in Solution, with the Domain for Dimerization in the N-Terminal Part of the Protein

Spaeny-Dekking

Hemert²,

P³

et al. 1995

Biochemistry

View full text Add to dashboard Cite

show abstract

“…If we assums that the elution profiles reflect the equilibria of the injected samples, the apparent dissociation constant [ K d(app) ] was estimated from fitting to eq . The injected samples were diluted during the chromatographic process; thus, the calculated K d(app) actually provides the upper limit of the K d . − The K d(app) values of the recombinant PZs are listed in Table . The K d(app) values of 113 ZL and 143 ZL in the dark state from our SEC analyses were 131 and 91 μM, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…The average molecular weight (MW) was determined using the calibration curve at the peak elution volume. The sample was assumed to be in a monomer–dimer equilibrium, and MW was described by the following equation: where M d is the molecular weight of the dimer calculated by the elution volume of each sample at 100 μM in the light state, M m is the molecular weight of the monomer (half of M d ), and α is the fraction of dimer as expressed by the following equation: α was plotted versus the injected concentration of the monomeric proteins ([monomer] inj ), and the apparent dissociation constant [ K d(app) ] was estimated from the fitting curve with the following equation: − …”

Section: Methodsmentioning

confidence: 99%

Molecular Mechanism of Photozipper, a Light-Regulated Dimerizing Module Consisting of the bZIP and LOV Domains of Aureochrome-1

Nakatani

Hisatomi

2015

Biochemistry

View full text Add to dashboard Cite

Aureochrome-1 (AUREO1) is a blue light (BL) receptor responsible for the BL-induced blanching of a stramenopile alga, Vaucheria frigida. The AUREO1 protein contains a central basic region/leucine zipper (bZIP) domain, and a C-terminal light-oxygen-voltage-sensing (LOV) domain. BL induces the dimerization of monomeric AUREO1, which subsequently increases the affinity of this transcription factor for its target DNA [Hisatomi, O., et al. (2014) J. Biol. Chem. 289, 17379-17391]. We constructed a synthetic gene encoding N-terminally truncated monomeric AUREO1 (designated Photozipper) to elucidate the molecular mechanism of this BL-regulated transcription factor and to develop it as an optogenetic tool. In this study, four different Photozipper (PZ) protein constructs were prepared comprising different N-terminal truncations. The monomer-dimer equilibria of the PZ constructs were investigated in the dark and light states. Dynamic light scattering and size-exclusion chromatography analyses revealed that the apparent dissociation constants of PZ dimers with and without the ZIP region were ~100 and 30 μM, respectively, indicating that the ZIP region stabilized the monomeric form in the dark state. In the light state, fluorescence resonance energy transfer analyses demonstrated that deletion of the ZIP region increased the dissociation constant from ~0.15 to 0.6 μM, suggesting that intermolecular LOV-LOV and ZIP-ZIP interactions stabilized the dimeric forms. Our results suggest that synergistic interactions between the LOV and bZIP domains stabilize the monomeric form in the dark state and the dimeric form in the light state, which possibly contributes to the function of PZ as a BL-regulated molecular switch.

show abstract

“…domain of y/ resolvase (43 amino acids) binds to res (or parts of res), but with a lower affinity than the intact protein [7]. Tn3 and y/ resolvases are dimeric in concentrated solution [12,13]. The dimer interface has been characterized crystallographically and by mutagenesis (reviewed in [5]).…”

Section: Introductionmentioning

confidence: 99%

Cooperative binding of Tn3 resolvase monomers to a functionally asymmetric binding site

et al. 1995

View full text Add to dashboard Cite

show abstract

Studies of the dimerization and domain structure of gamma delta resolvase

Cited by 13 publications

References 0 publications

Gin Invertase of Bacteriophage .mu. is a Dimer in Solution, with the Domain for Dimerization in the N-Terminal Part of the Protein

Gin Invertase of Bacteriophage .mu. is a Dimer in Solution, with the Domain for Dimerization in the N-Terminal Part of the Protein

Molecular Mechanism of Photozipper, a Light-Regulated Dimerizing Module Consisting of the bZIP and LOV Domains of Aureochrome-1

Cooperative binding of Tn3 resolvase monomers to a functionally asymmetric binding site

Contact Info

Product

Resources

About