The eukaryotic cofactor for the human immunodeficiency virus type 1 (HIV-1) rev protein, eIF-5A, maps to chromosome 17p12–p13: three eIF-5A pseudogenes map to 10q23.3, 17q25, and 19q13.2

The eukaryotic initiation factor 5A (eIF-5A) has been identified as an essential cofactor for the HIV-1 trans-activator protein Rev. Rev plays a key role in the complex regulation of HIV-1 gene expression and thereby in the generation of infectious virus particles. Expression of eIF-5A is vital for Rev function, and inhibition of this interaction leads to a block of the viral replication cycle. In humans, four different eIF-5A genes have been identified. One codes for the eIF-5A protein and the other three are pseudogenes. Using a panel of somatic rodent—human cell hybrids in combination with fluorescence in situ hybridization analysis, we show that the four genes map to threedifferent chromosomes. The coding eIF-5A gene (EIF5A) maps to 17p12–p13, and the three pseudogenes EIF5AP1, EIF5AP2, and EIF5AP3 map to 10q23.3, 17q25, and 19q13.2, respectively. This is the first localization report for a eukaryotic cofactor for a regulatory HIV-1 protein.     

An estimate of spin diffusion in a spin subset: Application to iterative distance calculation from 3D 15N NOESY-HMQC

Summary A method for quantification of distances between amide hydrogens using only the 3D NOESY-HMQC experiment recorded on a ¹⁵N-labelled protein is presented. This method is based on an approximate expression of the NOE intensities between amide hydrogens obtained from continuum modelling of the non-amide spins; this expression is used in a distance calculation algorithm. The algorithm has been named CROWD, standing for Continuum approximation of Relaxati On path Ways between Dilute spins. This approximation as well as the CROWD algorithm are tested on a simulated case; the CROWD algorithm is then applied to experimental data, measured on a fragment of bacteriorhodopsin.     

Three-dimensional structure of ectatomin from Ectatomma tuberculatum ant venom

Summary Two-dimensional ¹H NMR techniques were used to determine the spatial structure of ectatomin, a toxin from the venom of the ant Ectatomma tuberculatum. Nearly complete proton resonance assignments for two chains of ectatomin (37 and 34 amino acid residues, respectively) were obtained using 2D TOCSY, DQF-COSY and NOESY experiments. The cross-peak volumes in NOESY spectra were used to define the local structure of the protein and generate accurate proton-proton distance constraints employing the MARDIGRAS program. Disulfide bonds were located by analyzing the global fold of ectatomin, calculated with the distance geometry program DIANA. These data, combined with data on the rate of exchange of amide protons with deuterium, were used to obtain a final set of 20 structures by DIANA. These structures were refined by unrestrained energy minimization using the CHARMm program. The resulting rms deviations over 20 structures (excluding the mobile N- and C-termini of each chain) are 0.75 ? for backbone heavy atoms, and 1.25 ? for all heavy atoms. The conformations of the two chains are similar. Each chain consists of two α-helices and a hinge region of four residues; this forms a hairpin structure which is stabilized by disulfide bridges. The hinge regions of the two chains are connected together by a third disulfide bridge. Thus, ectatomin forms a four-α-helical bundle structure.     

Bioprocessing in space: Human cells attach to beads in microgravity

Attachment to a substrate and survival of human embryonic kidney (HEK) cells have been tested in an incubator installed in the flight-deck of the Space Shuttle ‘Challenger’ during its eighth mission.HEK cells are producing the enzyme urokinase and are presently investigated as candidates for electrophoretic separation in an apparatus developed and manufactured by McDonnell Douglas.Attachment of HEK cells to a substrate is mandatory for survival and production of urokinase after electrophoretic separation.Analysis of the samples shows that cells adhere, spread and survive in microgravity (< 10⁻³ ×g) conditions as well as the ground controls at 1 × g. This result represents an important step towards further bioprocessing in space.     

Enhancing Soybean Rhizosphere Colonization by Rhizobium japonicum

A study was conducted to seek means to increase the colonization of the rhizosphere of soybeans (Glycine max L. Merrill) by Rhizobium japonicum. For this purpose, a strain of R. japonicum that was resistant to benomyl, streptomycin, and erythromycin was used. The numbers of R. japonicum rose quickly in the first 2 days after soybean seeds were planted in soil and then rapidly fell. The decline was slower if the seeds were coated with benomyl. This fungicide reduced the numbers of bacteria and protozoa in the rhizosphere, but the effect became less or disappeared as the plants grew. In sterile soil inoculated with R. japonicum and a mixture of microorganisms, the numbers of R. japonicum were usually lower if protozoa were present than if they were absent. Nodulation and plant yield were increased by the addition of benomyl to soybean seeds sown in sterile soil inoculated with R. japonicum and a mixture of microorganisms. The addition of streptomycin and erythromycin to soil stimulated the growth of R. japonicum but inhibited other bacteria in the presence or absence of soybeans. The data indicate that colonization can be increased by the use of antimicrobial agents and R. japonicum strains resistant to those inhibitors.     

Physical mapping of cleavage sites recognized by restriction endonucleases on the genome of bacteriophage T5

The DNA of bacteriophage T5 has been treated with restriction endonucleases EcoRi, HindIII, BamI, SmaI, PstI, SalI, KpnI and the electrophoretic pattern obtained in agarose gel has been analyzed in order to localize the specific cleavage sites on the T5 DNA. The localization of cleavage sites has been deduced from the electrophoretic pattern of double and partial digests, the digests of isolated restriction fragments and the digests of deletion mutant T5st(o) DNA.Four BamI cleavage sites have been found and localized on the physical map of T5 DNA at 0.21, 0.225, 0.685 and 0.725 fractional length. Endonuclease SmaI cleaves at 0.39, 0.59 and 0.69 fractional length. Endonuclease PstI cuts T5 DNA at 11 sites: 0.090, 0.210, 0.320, 0.510, 0.635, 0.670, 0.705, 0.770, 0.815, 0.840, 0.875 fractional length. Six KpnI cleavage sites have been mapped at 0.170, 0.215, 0.525, 0.755, 0.830, 0.850 fractional length. A complete cleavage map of the phage genome is presented for seven restriction enzymes.     

New species ofAlchemilla ser.Hirsutae

The author describes four new apomict species ofAlchemilla L. ser.Hirsutae H. Lindb. They appear to be endemic in the Western Carpathians, growing there on some few mountain ranges, mostly in synanthropic habitats. Their known distribution areas seem to be ± separate. Two numerical indices called the epicalyx index and the hypanthium index respectively are defined to characterise the proportion of anAlchemilla flower. Ranges of both indices are shown for each of the new species.     

Observations on populations of the horseshoe crabLimulus (=Xiphosura) polyphemus

1. Marked populations of Limulus (=Xiphosura) polyphemus reveal that in Cold Spring Harbor, New York, they consisted of 10,000–18,000 adults in 1957 and 1961. The sex ratio in 1957 was about 4 males: 1 female. Pairs may remain attached for as long as 9 days. An undisturbed female may lay as many as 12,000 eggs in one nest.
2. The Cold Spring Harbor populations appear to be rather sedentary: none of the 1,000 animals marked on the north edge of the sandspit in 1961 were detected in the outer harbor either at Laurel Hollow Beach or the peninsula adjacent to the Cold Spring Harbor Yacht Club 500–800 meters from the tagging site (see Fig. 1), nor were they found in the small beach adjacent to the Biological Laboratory in the inner harbor. Similarly, none of the 300 animals marked at this last site were found at the north edge of the sandspit.
3. The phenotype of the compound eye varies from black to pigmentless. Samples observed in Cold Spring Harbor and in the Marine Biological Laboratory, Woods Hole, Massachusetts (separated by Long Island Sound and a distance of 150 miles) differ in the frequency of the various phenotypes scored, but the mode of inheritance of eye color remains obscure.
4. The available evidence indicates Limulus has considerable phenotypic variation in regard to body size, eye color, and other characters believed to be inherited, with the result that demes or physiological races are created. It is argued that the belief that this organism is stable and has not changed since the Triassic 200 million years ago has foundation only in regard to the pattern of the body of Limulus, but not in regard to its genotype. Limulus does not seem to be different from other organisms for which considerable genetic evidence is available, and thus the statement that DNA is fairly stable and has remained so for 200 million years is open to question.