Satellite DNA of the red flour beetle Tribolium castaneum--comparative study of satellites from the genus Tribolium

A highly abundant satellite DNA comprising 17% of the Tribolium castaneum (Insecta, Coleoptera) genome was cloned and sequenced. The satellite monomer is 360 bp long, has a high A+T content of 73%, and lacks significant internal substructures. The sequence variability is 3.6%, essentially due to random distribution of single-point mutations. The satellite is evenly distributed in the regions of centromeric heterochromatin of all 20 chromosomes, as shown by fluorescent in situ hybridization. Comparison of T. castaneum satellite with those from three different but congeneric species reveals the highest sequence similarity of 47.1% with the satellite from the sibling species Tribolium freemani. The phylogenetic relationships among Tribolium species deduced from satellite sequence agree with those based on karyological, chemotaxonomic, and hybridization data. This indicates a parallel in the divergence of satellites and some genetic and cytogenetic characters. Despite low mutual sequence similarity, which makes them species-specific, Tribolium satellites have a common structural characteristic: a block of about 95% A+T content, 20 to 42 bp long, flanked at one side by an inverted repeat which can potentially form a thermodynamically stable dyad structure. Since similar structural features are found in centromeric DNA of Saccharomyces cerevisiae and Chironomus pallidivittatus, their possible importance in centromere function may be inferred.      

Evidence for a direct interaction of Rev protein with nuclear envelop mRNA-translocation system.

The interaction of the Rev protein from human immunodeficiency virus type 1 (HIV-1) with the nucleocytoplasmic mRNA-transport system was investigated. In gel-shift assay, the recombinant Rev protein used in this study selectively bound to the Rev-responsive element (RRE) region of HIV-1 env-specific RNA. Nitrocellulose-filter-binding studies and Northern/Western-blotting experiments revealed an association constant of approximately 1 x 10(10) M-1. The Rev protein also strongly bound to isolated nuclear envelopes from H9 cells, containing the poly(A)-binding site (= mRNA carrier) and the nucleoside triphosphatase (= NTPase), which are thought to be involved in nuclear export of poly(A)-rich mRNA. Binding of 125I-Rev to a 110-kDa nuclear-envelope protein, the putative mRNA carrier, could be demonstrated in in vitro experiments. Both efflux of cellular poly(A)-rich RNA, such as actin RNA [but not efflux of poly(A)-free RNA] from isolated nuclei and the nuclear-envelope NTPase activity were strongly inhibited by Rev protein. On the other hand, transport of viral env RNA, containing the Rev-responsive element, was increased in the presence of Rev. Studying the release of RNA from closed nuclear-envelope vesicles containing entrapped RNA, the action of Rev was found to occur at the level of translocation of RNA through the nuclear pore. Evidence is presented that Rev down-regulates the NTPase-driven transport of mRNA lacking the RRE, most likely via binding to the mRNA carrier within the envelope. In contrast to the efflux of RRE-free RNA, ATP-dependent efflux of RRE-containing RNA from resealed nuclear-envelope vesicles was found to be increased, if the RNA was entrapped in the vesicles together with Rev protein. In addition, it was found that phosphorylated Rev, which is transported together with RRE-containing RNA out of the vesicles, becomes dephosphorylated during transport. In the vesicle experiments it is demonstrated for the first time that a protein selectively channels a specific mRNA across the nuclear-envelope pore complex.     

Development of anthropometric and physical performance profiles of young elite male soccer players: a longitudinal study

The purpose of the present longitudinal study was to explore distinctive anthropometric and physical performance characteristics of young soccer players between the age of 11 and 14 and to reveal the performance at the age of 11, which contributes to the later success. Male players of the best national male squads of the 'cadet league' (14 years of age; n = 26) were annually tested starting from the age of 11 for body size and composition, flexibility, power, coordination, and agility. Randomly selected untrained but physically active age-matched boys (n = 63) were also tested over 4 consecutive years. The results revealed no difference between 2 groups regarding the body size and composition (p > 0.05). The differences in flexibility emerged only at the later age, whereas the differences regarding the explosive power (as assessed by various jumps) were moderate and partly inconsistent. The most prominent advantage of the soccer players over the control subjects during the entire tested age period appeared to be movement agility and coordination (p < 0.01). Therefore, the explosive muscle power and, in particular, the agility and coordination characterize elite soccer players of 11-14 years of age but not the body size and body composition. In addition, the agility and coordination could be among the crucial factors of future success in 11-year-old players and, therefore, should be used for early selection.     

Complex structural features of satellite DNA sequences in the genus Pimelia (Coleoptera: Tenebrionidae): random differential amplification from a common 'satellite DNA library'

The major satellites of the nine species of the subgenera Pimelia s. str. and Amblyptera characterised in this paper are composed of longer monomers (500 and 700 bp) than those described previously in 26 Pimelia s. str. taxa (357 bp, a sequence called PIM357). Sequence analysis reveals partial similarity among these satellites and with the PIM357 monomers. The discrepancy between the phylogeny obtained based on three mitochondrial and two nuclear markers and that deduced from satellite DNA (stDNA) sequences suggests that the different Pimelia satellites were already present in a common ancestor forming what has been called a 'satellite DNA library'. Thus, the satellite profiles in the living species result from a random amplification of sequences from that 'library' during diversification of the species. However, species-specific turnover in the sequences has occurred at different rates. They have included abrupt replacements, a gradual divergence and, in other cases, no apparent change in sequence composition over a considerable evolutionary time. The results also suggest a common evolutionary origin of all these Pimelia satellite sequences, involving several rearrangements. We propose that the repeat unit of about 500 bp has originated from the insertion of a DNA fragment of 141 bp into the PIM357 unit. The 705-bp repeats have originated from a 32-bp direct duplication and the insertion of a 141-bp fragment in inverted orientation relative to a basic structure of 533 bp.     

Standard anthropometric, body composition, and strength variables as predictors of jumping performance in elite junior athletes

The aim of the study was to investigate whether variables routinely assessed while testing athletes can also predict movement performance. The relation between jumping performance and standard strength, anthropometric, and body composition variables was examined in elite junior basketball players. The 33 males were tested for maximal vertical jump, as well as for maximal isometric voluntary force and rate of force development of hip and knee extensors. Standard anthropometric and body composition measures (body height, lean body mass, as well as the percentage of fat and muscle tissue) were also taken. Except for maximal isometric forces (0.38 and 0.52 N.kg(-1) for hip and knee extensors, respectively), all correlation coefficients between the selected variables and jump height were insignificant. As a consequence, the corresponding multiple correlation coefficient, R = 0.71, also suggested a moderate predictability of jumping performance by the standard strength tests and anthropometric and body composition variables. The results obtained dispute the use of the examined tests in sport performance assessment, and also question applying the tests for other purposes such as evaluation of training procedures or selection of young athletes. Therefore, the results are in line with the concept that a reliable performance assessment in homogeneous groups of athletes requires predominantly movement-specific testing.     

Effect of maturation on the relationship between physical performance and body size

The purpose of the study was to explore the effect of maturation on the body size-physical performance relationship. Based on our previous study that evaluated only muscle strength, we hypothesized that the physical performance tested on pubescent subjects would be related to body size at a higher rate than predicted by standard scaling theory applied on pre- and postpubescent subjects. Six age groups of highly selected and trained junior soccer players (N = 478; age 12-17 years) presumably including prepubescent, pubescent, and postpubescent subjects were evaluated. They were tested using various standard tests of maximum physical performance including muscle strength of 3 leg muscle groups, 2 jumps, sit-ups, hand and foot tapping, and agility. The results revealed a steeper increase of the tested performance with an increase in body size for the pubescent age than for the pre- and postpubescent age. The observed phenomenon was interpreted by different level of maturation of the subjects tested within the same pubescent age group. We conclude that maturation alters the effect of body size on various physical performances and, therefore, this phenomenon needs to be taken into account when interpreting the data from the physical performance tests applied on pubescent subjects.     

Evolution of satellite DNAs from the genus Palorus--experimental evidence for the "library" hypothesis

Satellite DNA profiles have been characterized in the congeneric species Palorus ratzeburgii, Palorus subdepressus, Palorus genalis, and Palorus ficicola (Coleoptera, Insecta), each of which contains a single, A + T-rich satellite DNA comprising a considerable portion of the genome (20%-40%). These satellites exhibit insignificant mutual sequence similarity. Using PCR assay, it has been shown that all four sequences are present in each of the tested Palorus species: one of them is amplified into a high copy number or a major satellite, while the three others are in the form of low-copy-number repeats estimated to make up approximately 0.05% of the genome. Each of the four satellites is interspecifically high conserved concerning the sequence, monomer length, and tandem repeat organization. Major, as well as low- copy-number, satellites are colocalized in the regions of pericentromeric heterochromatin on all chromosomes of the complement. The low-copy-number satellites are dispersed between the large arrays of the major satellite over the whole heterochromatic block. Our results explain satellite DNA evolution, confirming the hypothesis that related species share a "library" of conserved satellite sequences, some of which could be amplified into a major satellite. Due to the evolutionary dynamics of satellite DNAs, the content of the "library" is variable; the elimination of some sequences parallels the creation of the new ones. Quantitative changes in satellite DNAs, induced by occasional amplification of satellite repeat from the "library", could possibly occur in the course of the speciation process, thus forming a species-specific profile of satellite DNAs.      

Evidence for involvement of a nuclear envelope-associated RNA helicase activity in nucleocytoplasmic RNA transport.

It seems well established that translocation of at least some mRNAs through the nuclear pore is (1) an energy-dependent process, and (2) dependent on the presence of the poly(A) segment attached to most mRNA species. We describe that RNA helicase (RNA duplex unwindase) activity is present in a nuclear envelope (NE) preparation, which also appears to be involved in nucleocytoplasmic RNA transport. This activity unwinds RNA: RNA hybrids. The helicase has a pH optimum of 7.5 and a temperature optimum of 30 degrees C. Applying the sealed NE vesicle system, it was shown that duplex RNA species are readily released from the vesicles in an unidirectional manner, in contrast to single-stranded RNA, which is much slower transported into the extravesicular space. Attachment of a poly(A) segment to the RNA duplex additionally increases the efflux rate of this RNA. Efflux of duplex RNA but not efflux of single-stranded RNA was strongly inhibited by formycin B 5'-triphosphate. Our results suggest that, besides poly(A), duplex structures, if present in a given RNA, modulate and control the export of RNA.     

Evaluation of the reliability of soccer-specific field tests

The soccer-specific field tests are popular among coaches due to their simplicity, validity, and minimal use of equipment. Nevertheless, there is a general lack of data about their reliability, particularly regarding the tests of anaerobic performance. Twenty professional male soccer players performed 3 consecutive trials of the tests of throwing-in and standing-kick performance (the distance measured) as well as on timed 10-m sprint, flying 20-m sprint, running 10 x 5 m, zigzag running with and without the ball, and the skill index (i.e., the ratio of the zigzag running without and with the ball). With the exception of the throwing-in and standing kick, the evaluated tests revealed high intraclass correlation coefficients (i.e., >0.80), small within-individual variations (coefficient of variation, <4%), and sample sizes for detecting a 2% change in the tested performance that are either close to or below the standard size of a professional soccer squad. In addition to simplicity and face validity, most of the evaluated tests revealed high reliability. Therefore, the evaluated tests are recommended for sport-specific profiling and early selection of young athletes as well as for routine testing procedures that could detect effects of various intervention procedures. Regarding the throwing-in and standing-kick tests, direct measurement of the ball velocity (e.g., with a standard radar gun) is recommended.