FISH mapping and molecular organization of the major repetitive sequences of tomato

This paper presents a bird’s-eye view of the major repeats and chromatin types of tomato. Using fluorescence in-situ hybridization (FISH) with Cot-1, Cot-10 and Cot-100 DNA as probes we mapped repetitive sequences of different complexity on pachytene complements. Cot-100 was found to cover all heterochromatin regions, and could be used to identify repeat-rich clones in BAC filter hybridization. Next we established the chromosomal locations of the tandem and dispersed repeats with respect to euchromatin, nucleolar organizer regions (NORs), heterochromatin, and centromeres. The tomato genomic repeats TGRII and TGRIII appeared to be major components of the pericentromeres, whereas the newly discovered TGRIV repeat was found mainly in the structural centromeres. The highly methylated NOR of chromosome 2 is rich in [GACA]₄, a microsatellite that also forms part of the pericentromeres, together with [GA]₈, [GATA]₄ and Ty1-copia. Based on the morphology of pachytene chromosomes and the distribution of repeats studied so far, we now propose six different chromatin classes for tomato: (1) euchromatin, (2) chromomeres, (3) distal heterochromatin and interstitial heterochromatic knobs, (4) pericentromere heterochromatin, (5) functional centromere heterochromatin and (6) nucleolar organizer region.     

Monoclonal antibodies against the 70-kilodalton iron-regulated protein of Neisseria meningitidis are bactericidal and strain specific.

When grown under iron limitation, Neisseria meningitidis expresses a number of outer membrane proteins (OMPs), one of which is a 70-kilodalton (kDa) major OMP. After immunization of mice with outer membrane preparations of iron-depleted cells of strain H44/76 (B:15:P1.7,16), hybridoma cell lines producing monoclonal antibodies against the 70-kDa OMP were obtained. Some of these monoclonal antibodies demonstrated strong bactericidal activity against the homologous strain H44/76 in the presence of human complement, suggesting potential application of the 70-kDa OMP as a vaccine component. However, none of the 10 selected monoclonal antibodies was able to recognize the corresponding protein from five heterologous strains of various serosubtyping characteristics. A polyclonal anti-70-kDa OMP serum also did not react with the other strains. This result shows that immunodominant surface-exposed epitopes of the meningococcal 70-kDa iron-limitation-inducible OMP are strain specific.     

Sequence comparison of human and yeast telomeres identifies structurally distinct subtelomeric domains

We have sequenced and compared DNA from the ends of three human chromosomes: 4p, 16p and 22q. In all cases the pro-terminal regions are subdivided by degenerate (TTAGGG)n repeats into distal and proximal sub- domains with entirely different patterns of homology to other chromosome ends. The distal regions contain numerous, short (<2 kb) segments of interrupted homology to many other human telomeric regions. The proximal regions show much longer (approximately 10-40 kb) uninterrupted homology to a few chromosome ends. A comparison of all yeast subtelomeric regions indicates that they too are subdivided by degenerate TTAGGG repeats into distal and proximal sub-domains with similarly different patterns of identity to other non-homologous chromosome ends. Sequence comparisons indicate that the distal and proximal sub-domains do not interact with each other and that they interact quite differently with the corresponding regions on other, non- homologous, chromosomes. These findings suggest that the degenerate TTAGGG repeats identify a previously unrecognized, evolutionarily conserved boundary between remarkably different subtelomeric domains.      

Molecular characterization and physical localization of highly repetitive DNA sequences from Brazilian Alstroemeria species

Highly repetitive DNA sequences were isolated from genomic DNA libraries of Alstroemeria psittacina and A. inodora. Among the repetitive sequences that were isolated, tandem repeats as well as dispersed repeats could be discerned. The tandem repeats belonged to a family of interlinked Sau3A subfragments with sizes varying from 68–127 bp, and constituted a larger HinfI repeat of approximately 400 bp. Southern hybridization showed a similar molecular organization of the tandem repeats in each of the Brazilian Alstroemeria species tested. None of the repeats hybridized with DNA from Chilean Alstroemeria species, which indicates that they are specific for the Brazilian species. In-situ localization studies revealed the tandem repeats to be localized in clusters on the chromosomes of A. inodora and A. psittacina: distal hybridization sites were found on chromosome arms 2PS, 6PL, 7PS, 7PL and 8PL, interstitial sites on chromosome arms 2PL, 3PL, 4PL and 5PL. The applicability of the tandem repeats for cytogenetic analysis of interspecific hybrids and their role in heterochromatin organization are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Facilitation and depression in the responses of spinal Renshaw cells to random stimulation of motor axons

1. We investigated the responses of cat lumbosacral Renshaw cells to pseudo-Poison stimulus sequences (of three different mean rates) delivered to motor axons in ventral roots or various muscle nerves. The Renshaw cell responses were evaluated by computation of peristimulus time histograms (PSTHs). 2. PSTHs computed with respect to all the stimuli showed, before the reference time, near-constant bin contents corresponding to the mean firing probability (rate), and an initial excitatory component (increase in discharge probability) after the reference time, followed by a small but longer-lasting reduction of firing rate. These two response components were strongly correlated linearly. It is suggested that the postexcitatory rate reduction is predominantly due to afterhyperpolarization. 3. In general, Renshaw cell responses to any stimulus in a stimulus train depended upon the stimulation history. In the averaged record, the response to the second of a pair of stimuli was affected by the first stimulus independently of intervening (random) stimuli. Very often, the second response showed a long-lasting depression (from 25 to greater than 250 ms). In a number of cases a briefer facilitating effect preceded the depression. 4. These conditioning effects were largely homosynaptic, i.e., confined to the particular input channel that was stimulated. This was shown by stimulating two different nerves (or nerve branches) with independent random patterns of similar mean rates and determining the cross-conditioning exerted by one input channel on the excitatory effects of the other. At small intervals between conditioning and test stimuli of some tens of milliseconds, a facilitatory effect could often be seen, which almost certainly reflected spatial summation. However, the subsequent depressant effect was largely accounted for by the postexcitatory rate reduction consequent to the conditioning stimulus in the parallel channel. Autoconditioning was still present. 5. The amount of facilitation and depression as well as their balance depended on the average Renshaw cell response. This in turn depended, at each mean stimulus rate, on the strength of synaptic coupling between an input channel and the cell, and on the mean stimulus rate, declining with an increase in mean rate. That is, the facilitation increased and the depression decreased with decreasing synaptic coupling and increasing mean stimulus rate. 6. Several factors may contribute to facilitation and depression; these are discussed with respect to their relative quantitative significance.(ABSTRACT TRUNCATED AT 400 WORDS)