An assessment of spatio-temporal genetic variation in the South African abalone (<Emphasis Type="Italic">Haliotis midae</Emphasis>), using SNPs: implications for conservation management

The South African abalone (Haliotis midae) is a gastropod mollusc of economic importance. In recent years natural populations have come under considerable pressure due to overharvesting and ecological shifts. The spatial genetic structure of H. midae has been determined; however there has not been a temporal assessment of abalone population dynamics around the South African coast. Using a population genomics approach this study aimed to assess fluctuations in genetic diversity among wild and cultured South African abalone populations through time and space. Various estimates of genetic diversity and population differentiation were calculated using EST-derived SNP markers. All populations had comparable levels of genetic diversity and the long-term effective population size appears to be sufficiently large for the wild populations, despite evidence of recent bottlenecks. Population differentiation was for the most part geographically correlated, with spatial genetic structure maintained across temporal samples. Significant genetic differentiation was, however, detected among temporal samples taken from the same locality. There was evidence for comparatively small short-term effective population sizes that could explain large changes in allele frequencies due to stochastic effects. Temporal heterogeneity could also be explained by changes in selection pressures over time. H. midae populations could, therefore, be more dynamic than previously estimated and this could have implications for effective conservation and fisheries management.     

Anatomy and ultrastructure of the marine oligochaete Tubificoides benedii (Tubificidae), with emphasis on its epidermis-cuticle-complex

Although Tubificoides benedii (d'Udekem, 1855) (= Peloscolex benedeni) is a ubiquitous form in eutrophicated or polluted coastal muds and is characterized by an exceptional viability in sulphide sediments, almost nothing is known about its anatomy and the structure of its conspicuously papillate body surface. As a part of a research project on sulphide annelids, done by our group, we investigated the body structure of this common and still extraordinary marine tubificid using light, scanning and transmission electron microscopy.While the internal structures correspond to the general pattern of marine tubificids (Giere, 1983), the epidermis-cuticle-complex is unusual. The epidermis cells, which are much interdigitated, contain numerous extremely long and irregularly shaped mitochondria. At the tip of the body, many sensory cells, embedded in the normal epidermis cell layer, end with ciliary tufts at the body surface. Long epidermal microvilli traverse the thick cuticular layer which only in the anterior- and posterior-most segments is studded by epicuticular projections. From most parts of the body these surficial rodlets, so far held typical for all Oligochaeta, are absent. Instead, here the cuticle forms numerous high, almost leaf-life papillae between which a viscous mucus layer regularly harbours many bacteria.This distinct papillate body armature may serve for stabilization of the mucus layer with its associated bacteria. It is well conceivable that the mucus, beside its usual role of reducting friction in the burrowing process, attracts bacteria. That their settlement may be more than an accidental association and involve some regulative interaction is indicated by the specific attachment of gram-negative bacterial threads often populating the posterior end of T. benedii (Dubilier, 1986). The significance of possible stress exerted by the sulfide-environment on the worm, as evidenced by an unusually high concentration of lysosomal structures and abnormally formed mitochondria in the epidermis, has to be verified in further studies.     

Characterization of the HeLa cell single-stranded DNA-dependent ATPase/DNA helicase II

A single-stranded DNA-dependent ATPase activity, consisting of two subunits of 83 kDa (p90) and 68 kDa (p70), was previously purified from HeLa cells (Vishwanatha, J.K. and Baril, E.F. (1990) Biochem 29, 8753–8759). Homology of the two subunits of single-stranded DNA-dependent ATPase with the human Ku protein (Caoet al. (1994) Biochem 33, 8548–8557) and identity of the Ku protein as the human DNA helicase II (Tutejaet al. (1994) EMBO J. 13, 4991–5001) have been reported recently. Using antisera raised against the subunits of the HDH II, we confirm that the Hela single-stranded DNA-dependent ATPase is the HDH II. Similar to the activity reported for Ku protein, ssDNA-dependent ATPase binds to double-stranded DNA and the DNA-protein complex detected by gel mobility shift assay consists of both the ATPase subunits. The p90 subunit is predominantly nuclear and is easily dissociated from chromatin. The p70 is distributed in cytosol and nucleus, and a fraction of the nuclear p70 protein is found to be associated with the nuclear matrix. Both the p90 and p70 subunits of the ATPase are present in G₁ and S phase of the cell cycle and are rapidly degraded in the G₂/M phase of the cell cycle.Abbreviations ssDNA single-stranded DNA - dsDNA double-stranded DNA - ATPase adenosine triphosphatase - HDH II human DNA helicase II - PGK 3-phosphoglycerate kinase     

Purification and characterization of proteins that bind to yeast ARSs

Two proteins that bind to yeast ARS DNA have been purified using conventional and oligonucleotide affinity chromatography. One protein has been purified to homogeneity and has a mass of 135 kDa. Competitive binding studies and DNase I footprinting show that the protein binds to a sequence about 80 base pairs away from the core consensus in the region known as domain B. This region has previously been shown to be required for efficient replication of plasmids carrying ARS1 elements. To investigate further whether the protein might have a function related to the ability of ARSs to act as replicators, binding to another ARS was tested. The protein binds to the functional ARS adjacent to the silent mating type locus HMR, called the HMR-E ARS, about 60 base pairs from the core consensus sequence. Surprisingly, there is little homology between the binding site at the HMR-E ARS and the binding site at ARS1. The 135-kDa protein is probably the same as ABF-I (SBF I) (Shore, D., Stillman, D. J. Brand, A. H., and Nasmyth, K. A. (1987) EMBO J. 6, 461-467; Buchman, A. R., Kimmerly, W. J., Rine, J., and Kornberg, R. D. (1988) Mol. Cell. Biol. 8, 210-225). A second DNA-binding protein was separated from ABF-I during later stages of the purification. This protein, which we designate ABF-III, also binds specifically to the ARS1 sequence, as shown by DNase I footprinting, at a site adjacent to the ABF-I recognition site. Purification of these two ARS binding proteins should aid in our understanding of the complex mechanisms that regulate eukaryotic DNA replication.