Crystal structure of a thermophilic GrpE protein: insight into thermosensing function for the DnaK chaperone system.

A homodimeric GrpE protein functions as a nucleotide exchange factor of the eubacterium DnaK molecular chaperone system. The co-chaperone GrpE accelerates ADP dissociation from, and promotes ATP binding to, DnaK, which cooperatively facilitates the DnaK chaperone cycle with another co-chaperone, DnaJ. GrpE characteristically undergoes two-step conformational changes in response to elevation of the environmental temperature. In the first transition at heat-shock temperatures, a fully reversible and functionally deficient structural alteration takes place in GrpE, and then the higher temperatures lead to the irreversible dissociation of the GrpE dimer into monomers as the second transition. GrpE is also thought to be a thermosensor of the DnaK system, since it is the only member of the DnaK system that changes its structure reversibly and loses its function at heat-shock temperatures of various organisms. We here report the crystal structure of GrpE from Thermus thermophilus HB8 (GrpE(Tth)) at 3.23 A resolution. The resolved structure is compared with that of GrpE from mesophilic Escherichia coli (GrpE(Eco)), revealing structural similarities, particularly in the DnaK interaction regions, and structural characteristics for the thermal stability of GrpE(Tth). In addition, the structure analysis raised the possibility that the polypeptide chain in the reported GrpE(Eco) structure was misinterpreted. Comparison of these two GrpE structures combined with the results of limited proteolysis experiments provides insight into the protein dynamics of GrpE(Tth) correlated with the shift of temperature, and also suggests that the localized and partial unfolding at the plausible DnaK interaction sites of GrpE(Tth) causes functional deficiency of nucleotide exchange factor in response to the heat shock

Negativity volume of the generalized Wigner function in gravitating hybrid system

We evaluate the gravity-induced negativity volume of the generalized Wigner function in a hybrid system consisting of a particle in a two-localized superposition state and an oscillator. The generalized Wigner function can capture the nonclassicality of the system. The increase in the negativity volume of the generalized Wigner function can be an indicator of the entanglement generation, which is demonstrated in the hybrid system generating the gravity-induced entanglement in various initial states. Moreover, by comparing the behaviors of the negativity volume with the entanglement fidelity, we show that the nonclassical feature of entanglement is properly identified by the criterion based on the negativity volume of the generalized Wigner function when the oscillator is initially in a thermal state.Comment: 12 pages, 5 figure

Racemic (RS C,SR S)-(2-{[1-allyl­oxy­carbonyl-3-(methyl­sulfanyl)prop­yl]iminometh­yl}phenyl-κ3 S,N,C 1)chlorido­platinum(II)

The title compound, [Pt(C15H18NO2S)Cl], was obtained by the cyclo­metallation reaction of cis-bis­(benzonitrile)dichlorido­platinum(II) with N-benzyl­idene-l-methio­nine allyl ester in refluxing toluene. The PtII atom has a square-planar geometry and is tetra-coordinated by the Cl atom and the C, N and S atoms from the benzyl­idene methio­nine ester ligand. In the crystal structure, the S atoms show opposite chiral configurations with respect to the α-carbon of the methio­nine, reducing steric repulsion between the methyl and allyl ester groups

Csm4-Dependent Telomere Movement on Nuclear Envelope Promotes Meiotic Recombination

During meiotic prophase, chromosomes display rapid movement, and their telomeres attach to the nuclear envelope and cluster to form a “chromosomal bouquet.” Little is known about the roles of the chromosome movement and telomere clustering in this phase. In budding yeast, telomere clustering is promoted by a meiosis-specific, telomere-binding protein, Ndj1. Here, we show that a meiosis-specific protein, Csm4, which forms a complex with Ndj1, facilitates bouquet formation. In the absence of Csm4, Ndj1-bound telomeres tether to nuclear envelopes but do not cluster, suggesting that telomere clustering in the meiotic prophase consists of at least two distinct steps: Ndj1-dependent tethering to the nuclear envelope and Csm4-dependent clustering/movement. Similar to Ndj1, Csm4 is required for several distinct steps during meiotic recombination. Our results suggest that Csm4 promotes efficient second-end capture of a double-strand break following a homology search, as well as resolution of the double-Holliday junction during crossover formation. We propose that chromosome movement and associated telomere dynamics at the nuclear envelope promotes the completion of key biochemical steps during meiotic recombination

Genetic Analysis of Baker's Yeast Msh4-Msh5 Reveals a Threshold Crossover Level for Meiotic Viability

During meiosis, the Msh4-Msh5 complex is thought to stabilize single-end invasion intermediates that form during early stages of recombination and subsequently bind to Holliday junctions to facilitate crossover formation. To analyze Msh4-Msh5 function, we mutagenized 57 residues in Saccharomyces cerevisiae Msh4 and Msh5 that are either conserved across all Msh4/5 family members or are specific to Msh4 and Msh5. The Msh5 subunit appeared more sensitive to mutagenesis. We identified msh4 and msh5 threshold (msh4/5-t) mutants that showed wild-type spore viability and crossover interference but displayed, compared to wild-type, up to a two-fold decrease in crossing over on large and medium sized chromosomes (XV, VII, VIII). Crossing over on a small chromosome, however, approached wild-type levels. The msh4/5-t mutants also displayed synaptonemal complex assembly defects. A triple mutant containing a msh4/5-t allele and mutations that decreased meiotic double-strand break levels (spo11-HA) and crossover interference (pch2Δ) showed synergistic defects in spore viability. Together these results indicate that the baker's yeast meiotic cell does not require the ∼90 crossovers maintained by crossover homeostasis to form viable spores. They also show that Pch2-mediated crossover interference is important to maintain meiotic viability when crossovers become limiting

Variations in amount of TSST-1 produced by clinical methicillin resistant Staphylococcus aureus (MRSA) isolates and allelic variation in accessory gene regulator (agr) locus

<p>Abstract</p> <p>Background</p> <p><it>Staphylococcus aureus </it>(S. aureus) is an important pathogen associated with both nosocomial and community-acquired infections and its pathogenicity is attributed to its potential to produce virulence factors. Since the amount of toxin produced is related to virulence, evaluating toxin production should be useful for controlling S. aureus infection. We previously found that some strains produce relatively large amounts of TSST-1; however, no reports have described the amount of TSST-1 produced by clinical isolates.</p> <p>Methods</p> <p>Amounts of TSST-1 produced by clinical methicillin resistant S. aureus (MRSA) isolates were measured by Western blotting. We determined their accessory gene regulator (<it>agr</it>) class by PCR and investigated whether TSST-1 production correlates with variations in the class and structure of the <it>agr</it>.</p> <p>Results</p> <p>We found that 75% of surveyed MRSA isolates (n = 152) possessed the <it>tst </it>gene and that 96.7% belonged to <it>agr </it>class 2. The concentrations of TSST-1 secreted into culture supernatants by 34 strains measured by Western blotting differed 170-fold. Sequencing the entire <it>agr </it>locus (n = 9) revealed that some had allelic variations regardless of the amount of TSST-1 produced whereas sequencing the <it>sar</it>, sigma factor B and the <it>tst </it>promoter region revealed no significant changes.</p> <p>Conclusion</p> <p>The amounts of TSST-1 produced by clinical MRSA isolates varied. The present results suggest that TSST-1 production is not directly associated with the <it>agr </it>structure, but is instead controlled by unknown transcriptional/translational regulatory systems, or synthesized by multiple regulatory mechanisms that are interlinked in a complex manner.</p