Strand separation establishes a sustained lock at the Tus-Ter replication fork barrier

The bidirectional replication of a circular chromosome by many bacteria necessitates proper termination to avoid the head-on collision of the opposing replisomes. In Escherichia coli, replisome progression beyond the termination site is prevented by Tus proteins bound to asymmetric Ter sites. Structural evidence indicates that strand separation on the blocking (nonpermissive) side of Tus-Ter triggers roadblock formation, but biochemical evidence also suggests roles for protein-protein interactions. Here DNA unzipping experiments demonstrate that nonpermissively oriented Tus-Ter forms a tight lock in the absence of replicative proteins, whereas permissively oriented Tus-Ter allows nearly unhindered strand separation. Quantifying the lock strength reveals the existence of several intermediate lock states that are impacted by mutations in the lock domain but not by mutations in the DNA-binding domain. Lock formation is highly specific and exceeds reported in vivo efficiencies. We postulate that protein-protein interactions may actually hinder, rather than promote, proper lock formation.Accepted Author ManuscriptBN/Nynke Dekker LabBN/Cees Dekker LabBN/Martin Depken La

Preparation of 3-Sulfonylated 3,3-Disubstituted Oxindoles by the Addition of Sulfinate Salts to 3-Halooxindoles

An efficient method for the preparation of 3-sulfonylated 3,3-disubstituted oxindole derivatives has been developed. The protocol involves a base-catalyzed addition of sulfinate salts to 3-halooxindoles, affording a wide range of 3-sulfonylated 3,3-disubstituted oxindoles in good to excellent yields under mild conditions. A preliminary trial of asymmetric catalytic version was conducted and gave promising enantioselectivity. The mechanism for the reaction was tentatively explored with the help of mass spectrometric analysis

Asymmetric Michael/Cyclization Cascade Reaction of 3-Isothiocyanato Oxindoles and 3-Nitroindoles with Amino-Thiocarbamate Catalysts: Enantioselective Synthesis of Polycyclic Spirooxindoles

An unprecedented organocatalytic asymmetric Michael/cyclization cascade reaction of 3-isothiocyanato oxindoles and. 3-nitroindoles has-been disclosed. A wide range of enantioenriched polycyclic, spirooxindoles, containing three contiguous chiral centers With two of them having quaternary stereocenters, could be smoothly obtained with satisfactory results (up to 99%-yield, >99:1 dr, and 96% ee). This methods very promising because the reaction is scalable, and the versatile transformations of the products into other spirocyclic oxindoles are also feasible