Letter to the Editor: 1H, 15N, and 13C chemical shift assignments of the resuscitation promoting factor domain of Rv1009 from Mycobacterium tuberculosis

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Détermination structurale de la cassiicoline, la toxine glycosylée de Corynespora cassiicola

Corynespora cassiicola est un champignon phytopathogène nécrotrophe capable d'infecter plus de 70 plantes hôtes, parmi lesquelles, la tomate, le concombre, le coton, le soja ou l'hévéa. Dans le cas de l'hévéa, la pathologie induite par C.!cassiicola se manifeste par des défoliations massives pouvant conduire à la mort de l'arbre. Cette infection est en constante progression dans toutes les zones de production

Pseudorandom Selective Excitation in NMR

In this work, average Hamiltonian theory is used to study selective excitation in a spin-1/2 system evolving under a series of small flip-angle $\theta-$pulses $(\theta\ll 1)$ that are applied either periodically [which corresponds to the DANTE pulse sequence] or aperiodically. First, an average Hamiltonian description of the DANTE pulse sequence is developed; such a description is determined to be valid either at or very far from the DANTE resonance frequencies, which are simply integer multiples of the inverse of the interpulse delay. For aperiodic excitation schemes where the interpulse delays are chosen pseudorandomly, a single resonance can be selectively excited if the $\theta$-pulses' phases are modulated in concert with the time delays. Such a selective pulse is termed a pseudorandom-DANTE or p-DANTE sequence, and the conditions in which an average Hamiltonian description of p-DANTE is found to be similar to that found for the DANTE sequence. It is also shown that averaging over different p-DANTE sequences that are selective for the same resonance can help reduce excitations at frequencies away from the resonance frequency, thereby improving the apparent selectivity of the p-DANTE sequences. Finally, experimental demonstrations of p-DANTE sequences and comparisons with theory are presented.Comment: 23 pages, 8 figure

The structure of a resuscitation-promoting factor domain from Mycobacterium tuberculosis shows homology to lysozymes

Resuscitation-promoting factor (RPF) proteins reactivate stationary-phase cultures of (G+C)-rich Gram-positive bacteria including the causative agent of tuberculosis, Mycobacterium tuberculosis. We report the solution structure of the RPF domain from M. tuberculosis Rv1009 (RpfB) solved by heteronuclear multidimensional NMR. Structural homology with various glycoside hydrolases suggested that RpfB cleaved oligosaccharides. Biochemical studies indicate that a conserved active site glutamate is important for resuscitation activity. These data, as well as the presence of a clear binding pocket for a large molecule, indicate that oligosaccharide cleavage is probably the signal for revival from dormancy

Protein kinase B controls Mycobacterium tuberculosis growth via phosphorylation of the transcriptional regulator Lsr2 at threonine 112.

Mycobacterium tuberculosis (Mtb) is able to persist in the body through months of multi-drug therapy. Mycobacteria possess a wide range of regulatory proteins, including the protein kinase B (PknB) which controls peptidoglycan biosynthesis during growth. Here, we observed that depletion of PknB resulted in specific transcriptional changes that are likely caused by reduced phosphorylation of the H-NS-like regulator Lsr2 at threonine 112. The activity of PknB towards this phosphosite was confirmed with purified proteins, and this site was required for adaptation of Mtb to hypoxic conditions, and growth on solid media. Like H-NS, Lsr2 binds DNA in sequence-dependent and non-specific modes. PknB phosphorylation of Lsr2 reduced DNA binding, measured by fluorescence anisotropy and electrophoretic mobility shift assays, and our NMR structure of phosphomimetic T112D Lsr2 suggests that this may be due to increased dynamics of the DNA-binding domain. Conversely, the phosphoablative T112A Lsr2 had increased binding to certain DNA sites in ChIP-sequencing, and Mtb containing this variant showed transcriptional changes that correspond with the change in DNA binding. In summary, PknB controls Mtb growth and adaptations to the changing host environment by phosphorylating the global transcriptional regulator Lsr2

Letter to the Editor: 1 H, 15 N and 13 C chemical shift assignments of the Pleckstrin Homology domain of the Human Protein Kinase B (PKB/Akt)

International audienc

Solution Structure of Human p8 MTCP1 , a Cysteine-rich Protein Encoded by the MTCP1 Oncogene, Reveals a New a a a-Helical Assembly Motif

International audienceMature-T-Cell Proliferation) is the ®rst gene unequivocally identi®ed in the group of uncommon leukemias with a mature phenotype. The three-dimensional solution structure of the human p8 MTCP1 protein encoded by the MTCP1 oncogene was determined by homonuc-lear proton two-dimensional NMR methods at 600 MHz. After sequence speci®c assignments, a total of 931 distance restraints and 57 dihedral restraints were collected. The location of the three previously unassigned disul®de bridges was determined from preliminary DIANA structures, using a statistical analysis of intercystinyl distances. The solution structure of p8 MTCP1 is presented as a set of 30 DIANA structures, further re®ned by restrained molecular dynamics using a simulated annealing protocol with the AMBER force ®eld. The r.m.s.d. values with respect to the mean structure for the backbone and all heavy atoms for a family of 30 structures are 0.73(AE0.28) and 1.17(AE0.23) A Ê , when the structured core of the protein (residues 5 to 63) is considered. The solution structure of p8 MTCP1 reveals an original scaffold consisting of three a helices, associated with a new cysteine motif. Two of the helices are covalently paired by two disul®de bridges, forming an a-hairpin which resembles an antiparallel coiled-coil. The third helix is oriented roughly parallel to the plane de®ned by the a-antiparallel motif and its axis forms an angle of %60 with respect to the main axis of this motif