Isolation and Characterization of Staphylococcus aureus Mutants Which Form Altered Cell Clusters

Staphylococcus aureus FDA 209P produces two extracellular bacteriolytic enzymes, 51-kDa endo-β-N-acetylglucosaminidase (GL) and 62-kDa N-acetylmuramyl-l -alanine amidase (AM), both of which can disperse cell clusters. To characterize the physiological roles of these enzymes in vivo, mutants with altered autolysin activity were isolated, and their degree of cluster formation in broth culture was assessed. Bacteriolytic activities of GL and AM, produced and secreted from these mutants into the culture fluid and detected with activity gels, coincided well with the degree of cluster formation of the mutants. The mutants with little or no enzyme activity grew in clusters, whereas those with high activity grew as well-separated cocci, suggesting that these enzymes are involved in cell separation of S. aureus in vivo.     

Mitogenomic evaluation of the historical biogeography of cichlids toward reliable dating of teleostean divergences

Background  

Recent advances in DNA sequencing and computation offer the opportunity for reliable estimates of divergence times between organisms based on molecular data. Bayesian estimations of divergence times that do not assume the molecular clock use time constraints at multiple nodes, usually based on the fossil records, as major boundary conditions. However, the fossil records of bony fishes may not adequately provide effective time constraints at multiple nodes. We explored an alternative source of time constraints in teleostean phylogeny by evaluating a biogeographic hypothesis concerning freshwater fishes from the family Cichlidae (Perciformes: Labroidei).     

Macrophages containing langerhans cell granules in normal lymph nodes of the rabbit

Summary Light and electron microscopic studies on macrophages of normal rabbit lymph nodes showed two types, one with little phagocytic activity and many features similar to those of epidermal Langerhans cells. Among these are characteristic Langerhans cell granules. From these findings it is concluded that the Langerhans cells may be derived from lymph node macrophages.The helpful advice and criticism of Dr. Toshio Nagano, Department of Anatomy, are gratefully acknowledged. The discussion with Dr. Mitsumasa Itoh, Department of Dermatology, was also helpful.     

Transport-coupled ubiquitination of the borate transporter BOR1 for its boron-dependent degradation

Plants take up and translocate nutrients through transporters. In Arabidopsis thaliana, the borate exporter BOR1 acts as a key transporter under boron (B) limitation in the soil. Upon sufficient-B supply, BOR1 undergoes ubiquitination and is transported to the vacuole for degradation, to avoid overaccumulation of B. However, the mechanisms underlying B-sensing and ubiquitination of BOR1 are unknown. In this study, we confirmed the lysine-590 residue in the C-terminal cytosolic region of BOR1 as the direct ubiquitination site and showed that BOR1 undergoes K63-linked polyubiquitination. A forward genetic screen identified that amino acid residues located in vicinity of the substrate-binding pocket of BOR1 are essential for the vacuolar sorting. BOR1 variants that lack B-transport activity showed a significant reduction of polyubiquitination and subsequent vacuolar sorting. Coexpression of wild-type (WT) and a transport-defective variant of BOR1 in the same cells showed degradation of the WT but not the variant upon sufficient-B supply. These findings suggest that polyubiquitination of BOR1 relies on its conformational transition during the transport cycle. We propose a model in which BOR1, as a B transceptor, directly senses the B concentration and promotes its own polyubiquitination and vacuolar sorting for quick and precise maintenance of B homeostasis.

The borate transporter BOR1 senses the boron concentration through its borate transport activity for K63-linked polyubiquitination and subsequent degradation.     

Countercurrent chromatography with a new horizontal flow-through coil planet centrifuge

A versatile horizontal flow-through coil planet centrifuge has been introduced for performing countercurrent chromatography. The apparatus carries a pair of coiled separation columns. Both of these allow continuous elution simultaneously without the use of rotating seals. One column enables preparative-scale separations and the other, analytical-scale separations, both with a high partition efficiency comparable to that obtained in liquid chromatography but without the complications arising from the use of solid supports. Capability of the apparatus was demonstrated on separation of dinitrophenyl amino acids using a two-phase solvent system composed of CHCl₃/CH₃COOH/0.1 HCl at a 2/2/1 volume ratio.     

Dynamic property of intermediate filaments: Regulation by phosphorylation

Site-specific phosphorylation of intermediate filament (IF) proteins on serine and threonine residues leads to alteration of the filament structure, in vitro and in vivo. Protein kinases involved in cell signaling and those activated in mitosis dynamically control spatial and temporal organization of intracellular IF phosphorylation. Thus, IF phosphorylation appears to be one of the most predominant strategies in coordinating intracellular organization of the IF network.