The BOS1 gene encodes an essential 27-kD putative membrane protein that is required for vesicular transport from the ER to the Golgi complex in yeast

We recently described the identification of BOS1 (Newman, A., J. Shim, and S. Ferro-Novick. 1990. Mol. Cell. Biol. 10:3405-3414.). BOS1 is a gene that in multiple copy suppresses the growth and secretion defect of bet1 and sec22, two mutants that disrupt transport from the ER to the Golgi complex in yeast. The ability of BOS1 to specifically suppress mutants blocked at a particular stage of the secretory pathway suggested that this gene encodes a protein that functions in this process. The experiments presented in this study support this hypothesis. Specifically, the BOS1 gene was found to be essential for cellular growth. Furthermore, cells depleted of the Bos1 protein fail to transport pro-alpha-factor and carboxypeptidase Y (CPY) to the Golgi apparatus. This defect in export leads to the accumulation of an extensive network of ER and small vesicles. DNA sequence analysis predicts that Bos1 is a 27-kD protein containing a putative membrane-spanning domain. This prediction is supported by differential centrifugation experiments. Thus, Bos1 appears to be a membrane protein that functions in conjunction with Bet1 and Sec22 to facilitate the transport of proteins at a step subsequent to translocation into the ER but before entry into the Golgi apparatus.     

Biomedicinals from the phytosymbionts of marine invertebrates: a molecular approach

Marine invertebrate animals such as sponges, gorgonians, tunicates and bryozoans are sources of biomedicinally relevant natural products, a small but growing number of which are advancing through clinical trials. Most metazoan and anthozoan species harbour commensal microorganisms that include prokaryotic bacteria, cyanobacteria (blue-green algae), eukaryotic microalgae, and fungi within host tissues where they reside as extra- and intra-cellular symbionts. In some sponges these associated microbes may constitute as much as 40% of the holobiont volume. There is now abundant evidence to suggest that a significant portion of the bioactive metabolites thought originally to be products of the source animal are often synthesized by their symbiotic microbiota. Several anti-cancer metabolites from marine sponges that have progressed to pre-clinical or clinical-trial phases, such as discodermolide, halichondrin B and bryostatin 1, are thought to be products derived from their microbiotic consortia. Freshwater and marine cyanobacteria are well recognised for producing numerous and structurally diverse bioactive and cytotoxic secondary metabolites suited to drug discovery. Sea sponges often contain dominant taxa-specific populations of cyanobacteria, and it is these phytosymbionts (= photosymbionts) that are considered to be the true biogenic source of a number of pharmacologically active polyketides and nonribosomally synthesized peptides produced within the sponge. Accordingly, new collections can be pre-screened in the field for the presence of phytobionts and, together with metagenomic screening using degenerate PCR primers to identify key polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) genes, afford a biodiscovery rationale based on the therapeutic prospects of phytochemical selection. Additionally, new cloning and biosynthetic expression strategies may provide a sustainable method for the supply of new pharmaceuticals derived from the uncultured phytosymbionts of marine organisms.     

Recombination and oligonucleotide analysis of guanidine-resistant foot-and-mouth disease virus mutants.

Guanidine resistance (gr) mutations of foot-and-mouth disease virus were mapped by recombining pairs of temperature-sensitive mutants belonging to different subtypes. In each cross, one parent possessed a gr mutation. Recombinants were isolated by selection at the nonpermissive temperature and assayed for the ability to grow in the presence of guanidine. From the progeny of three crosses, four different types of recombinant were distinguished on the basis of protein composition and RNA fingerprint. The sequences of the RNase T1-resistant oligonucleotides were determined and located in the full-length sequence of foot-and-mouth disease virus. The resulting maps show that (i) each recombinant was generated by a single genetic crossover, and (ii) both of the gr mutations studied were located within an internal 2.9-kilobase region which spans the P34 gene. This supports our hypothesis that guanidine inhibits the growth of foot-and-mouth disease virus by acting on nonstructural polypeptide P34. Additional evidence was provided by RNA fingerprinting gr mutants. In two of four cases the gr mutation was associated with a change in an oligonucleotide located near the 3' end of the P34 gene; in one of these the nucleotide substitution was identified.     

A New Naked Jawless Vertebrate From The Middle Devonian Of Scotland

A new jawless vertebrate, Achanarella trewini gen. et sp. nov., is described from some recently discovered, relatively well–preserved specimens from the Achanarras fish bed at Achanarras Quarry, northern Scotland. The Achanarras fish bed is of Middle Devonian (Eifelian) age. Achanarella is a torpedo–shaped jawless vertebrate with a strongly hypocercal tail and well–developed anal fin. It has at least 13 branchial openings with possibly more than 20. It is scaleless. Achanarella compares well with the Frasnian Euphaneropidae of the Escuminac Formation in Quebec, Canada, and Cornovichthys blaauweni Newman and Trewin, 2001, also from the Achanarras fish bed. It differs from these forms mainly in being extremely long and thin and having a very small head compared with body length.     

Nitrate supply affects ammonium transport in canola roots

Plants may suffer from ammonium (NH4+) toxicity when NH4+ is the sole nitrogen source. Nitrate (NO3-) is known to alleviate NH4+ toxicity, but the mechanisms are unknown. This study has evaluated possible mechanisms of NO3- alleviation of NH4+ toxicity in canola (Brassica napus L.). Dynamics of net fluxes of NH4+, H+, K+ and Ca2+ were assessed, using a non-invasive microelectrode (MIFE) technique, in plants having different NO3- supplies, after single or several subsequent increases in external NH4Cl concentration. After an increase in external NH4Cl without NO3-, NH4+ net fluxes demonstrated three distinct stages: release (tau1), return to uptake (tau2), and a decrease in uptake rate (tau3). The presence of NO3- in the bathing medium prevented the tau1 release and also resulted in slower activation of the tau3 stage. Net fluxes of Ca2+ were in the opposite direction to NH4+ net fluxes, regardless of NO3- supply. In contrast, H+ and K+ net fluxes and change in external pH were not correlated with NH4+ net fluxes. It is concluded that (i) NO3- primarily affects the NH4+ low-affinity influx system; and (ii) NH4+ transport is inversely linked to Ca2+ net flux.     

Morphological and allozyme diversity in the Hieracium nigrescens group (Compositae) in the Sudety Mountains and the Western Carpathians

The overall pattern of morphological variation and genetic diversity (allozyme analysis) was studied in the Hieracium nigrescens group (H. nigrescens s.l., H. alpinum ≥ H. murorum) in the Sudety Mountains and the Western Carpathians. A morphological analysis was performed on 180 plants from 12 populations belonging to six a priori distinguished taxa. Altogether, 25 characters were measured or scored. Morphometric (canonical discriminant analysis) data separated five taxa, evaluated here at the species rank: H. chrysostyloides, H. decipiens, H. nigrescens (all from the Sudety Mountains), H. jarzabczynum, and H. vapenicanum (the Western Carpathians). A distinct local population from Mount Babia hora (the Western Carpathians) comprised a further possible taxon, given the preliminary name ‘H. babiagorense’. Genetic diversity was studied in 17 populations of H. chrysostyloides, H. decipiens, H. jarzabczynum, H. nigrescens, H. vapenicanum and ‘H. babiagorense’ using five enzyme systems. All a priori recognized species were proved to be genetically homogeneous, each consisting of one unique multilocus allozyme genotype, except ‘H. babiagorense’ which shared the same genotype with H. jarzabczynum. For the first time, a chromosome number is reported for H. vapenicanum (2n = 3x = 27) and previously published numbers were confirmed for H. chrysostyloides (2n = 5x = 45), H. decipiens (2n = 4x = 36), H. jarzabczynum (2n = 4x = 36), H. koprovanum (2n = 4x = 36), and H. nigrescens (2n = 4x = 36). All species have been shown to be endemic to either the Sudety Mountains or the Western Carpathians. Except for the species studied, two further ones (H. apiculatum, H. nivimontis) are recognized in the area, giving a total of seven species from the Hieracium nigrescens group in the area studied. The morphologically slightly different local population from Mount Babia hora/Babia Góra (‘H. babiagorense’) requires further study. Two new combinations are proposed: Hieracium jarzabczynum (Paw?. & Zahn) Mráz & Chrtek f. and Hieracium vapenicanum (Lengyel & Zahn) Chrtek f. & Mráz. © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society, 2007, 153 , 287–300.