Protein-protein interaction in transport

The transport of histidine in the gram negative bacterium S. typhimurium has been studied over a number of years and found to occur through five transport systems (Ames, 1972). Of these, the one with the highest affinity has been studied in detail from the genetic, physiological and biochemical point of view. This system, known as the high-affinity histidine permease, is composed of two subsystems, the J-P and K-P systems, which have a component in common, the P protein, presumed to be membrane-bound. The J-P system, moreover, is known to require the presence of a periplasmic histidine-binding protein, the J protein. The J protein is coded for by the hisJ gene and the P protein is coded for by the hisP gene. Both of these genes have been mapped at 75 min on the Salmonella chromosomal map. Adjacent to them is a regulatory gene, the dhuA gene. The periplasmic histidine-binding protein J has been shown to interact directly with the second component of transport, the P protein (Ames and Spudich, 1976). In accordance with this, histidine-binding protein J has been shown to contain, besides the histidine-binding site, a second site, essential for function, the interaction site (Kustu and Ames, 1974). We have recently shown that a mutant J protein with a defective interaction site but an intact histidine-binding site cannot function in histidine transport, unless an appropriate compensating mutation is introduced in the P protein. The interaction between the J and P proteins is an obligatory step in transport. The mutation in the interaction site of the J protein has been shown to map in the hisJ gene, and the compensating supressor mutation in the P protein has been shown to map in the hisP gene. Our contention that the J and P proteins engage in a functional interaction assumes further strength from other studies on protein-protein interaction in bacteriophage development and in ribosomal structure. Among the possible functions of the J-P interaction in histidine transport, a likely one is the transmission of information to the P protein, concerning whether or not the histidine-binding site on the J protein is occupied. Appropriate conformational changes then can occur in either the J or the P protein, or both, such that the histidine is released in the correct location and direction on the inside of the cell. This could occur either by a pore-formation mechanism or by binding-site translocation. Another alternative is that the P protein is part of an energy transducing mechanism in which energy is transmitted to the J protein, through the interaction site, as a prerequisite for the J protein participation in translocation. Among the interesting findings coming out of this work, is also the fact that the P protein performs a central function in transport being involved in the permeation of other substrates besides histidine. It is likely that other binding proteins besides the J protein require the P protein. Thus an interesting question which we are trying to answer at present is whether the P protein has separate interaction sites for each of these other binding proteins requiring its function, or whether they all interact at one common site.     

Clonal descent and microevolution of Neisseria meningitidis during 30 years of epidemic spread

Serogroup A meningococci of subgroups III, IV-1 and IV-2 are probably descended from a common ancestor that existed in the nineteenth century. The 10.5 kb sequences spanning five distinct chromosomal loci, encoding cell-surface antigens, a secreted protease or housekeeping genes and intergenic regions, were almost identical in strains of those subgroups isolated in 1966, 1966 and 1917 respectively. During the subsequent two to three decades, all of these loci varied as a result of mutation, translocation or import of DNA from unrelated neisseriae. Thus, microevolution occurs frequently in naturally transformable bacteria. Many variants were isolated only once or within a single geographical location and disappeared thereafter. Other variants achieved genetic fixation within months or a few years. The speed with which sequence variation is either eliminated or fixed may reflect sequential bottlenecks associated with epidemic spread and contrasts with the results of phylogenetic analyses from bacteria that do not cause epidemics.     

Il genere Allium L. in Italia. IV. Studio citofotometrico sul granulo pollinico di Allium Chamaemoly L.

Abstract

The genus Allium L. in Italy. IV. A DNA cytophotometric study on the pollen grain of Allium chamaemoly L. — A cytophotometric analysis of DNA contents in pollen generative and vegetative nuclei of Allium chamaemoly L. was carried out. DNA synthesis in both nuclei was confirmed and a lightly higher DNA amount than 2C in the vegetative nucleus was pointed out. An analysis of the Fast-green stainable histones in the generative and vegetative nuclei was also accomplished. While the generative nucleus had a very high content of Fast-green stainable histone, the vegetative one have nearly no stainable histone. The occurrence of DNA synthesis and the very low histone content suggest the vegetative nucleus is functional and biochemically activ. The higher than 2C DNA content supports the possibility of a DNA amplification process including probably the amplification of ribosomal cistrons in the pollen vegetative nucleus.     

Acyl-CoA: sn-Glycerol-3-Phosphate O-Acyltransferase in Rat Brain Mitochondria and Microsomes

Abstract: The subcellular distribution of acyl-CoA: sn -glycerol-3-phosphate O-acyltransferase between brain mitochondria and microsomes was investigated. The activities associated with purified rat brain mitochondrial and microsomal preparations could be distinguished by differences in their acyl-CoA specificity, products of acylation, and sensitivity to N -ethylmaleimide, trypsin, acetone, and polymyxin B. It was concluded that both brain mitochondria and microsomes possess the acyltransferase.     

A phloroglucinol derivative from the brown alga Zonaria tournefortii

The major lipid component of the brown seaweed Zonaria tournefortii was identified as the acylphlorogluconol (all Z)-2′-icosa-5,8,11,14,17-pentae     

Nusinersen treatment and cerebrospinal fluid neurofilaments: An explorative study on Spinal Muscular Atrophy type 3 patients

The antisense oligonucleotide Nusinersen has been recently licensed to treat spinal muscular atrophy (SMA). Since SMA type 3 is characterized by variable phenotype and milder progression, biomarkers of early treatment response are urgently needed. We investigated the cerebrospinal fluid (CSF) concentration of neurofilaments in SMA type 3 patients treated with Nusinersen as a potential biomarker of treatment efficacy. The concentration of phosphorylated neurofilaments heavy chain (pNfH) and light chain (NfL) in the CSF of SMA type 3 patients was evaluated before and after six months since the first Nusinersen administration, performed with commercially available enzyme-linked immunosorbent assay (ELISA) kits. Clinical evaluation of SMA patients was performed with standardized motor function scales. Baseline neurofilament levels in patients were comparable to controls, but significantly decreased after six months of treatment, while motor functions were only marginally ameliorated. No significant correlation was observed between the change in motor functions and that of neurofilaments over time. The reduction of neurofilament levels suggests a possible early biochemical effect of treatment on axonal degeneration, which may precede changes in motor performance. Our study mandates further investigations to assess neurofilaments as a marker of treatment response.     

Inhibition of bromodomain and extra‐terminal proteins increases sensitivity to venetoclax in chronic lymphocytic leukaemia

The development of drugs able to target BTK, PI3k‐delta and BCL2 has dramatically improved chronic lymphocytic leukaemia (CLL) therapies. However, drug resistance to these therapies has already been reported due to non‐recurrent changes in oncogenic pathways and genes expression signatures. In this study, we investigated the cooperative role of the BCL2 inhibitor venetoclax and the BRD4 inhibitor JQ1. In particular, we found that JQ1 shows additional activity with venetoclax, in CLL cell lines and in ex vivo isolated primary CD19⁺ lymphocytes, arguing in favour of combination strategies. Lastly, JQ1 is also effective in venetoclax‐resistant CLL cell lines. Together, our findings indicated that the BET inhibitor JQ1 could be a promising therapy in CLL, both as first‐line therapy in combination with venetoclax and as second‐line therapy, after the emergence of venetoclax‐resistant clones.     

Antiproliferative Effect and Cell Cycle Alterations Induced by Salvia officinalis Essential Oil and Its Three Main Components in Human Colon Cancer Cell Lines

Colon cancer is one of the most common human malignancies, and chemotherapy cannot yet prevent recurrence in all patients. Essential oils are phytocomplexes with antiproliferative properties. In this study, we elucidated the antiproliferative properties and the effect on cell cycle progression of Sicilian Salvia officinalis essential oil and its three main compounds, α‐thujone, 1,8‐cineole (eucalyptol) and camphor, on three human colon cancer cell lines. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography. Cell proliferation was evaluated by MTT assay, and the cell cycle distribution was determined by flow cytometry. Thirty‐four compounds were identified in the tested essential oil. Growth inhibition was observed after 72 h, with an impact on cell cycle progression and no effect on the viability of normal colonic epithelial cells. The study shows that S. officinalis essential oil and its three main components have an in vitro antiproliferative effect on colon cancer cells.