Subcellular localization of chlorosome proteins in Chlorobium tepidum and characterization of three new chlorosome proteins: CsmF, CsmH, and CsmX

Chlorosomes are unique light-harvesting structures found in two families of photosynthetic bacteria. In this study, three chlorosome proteins (CsmF, CsmH, and CsmX) of the green sulfur bacterium Chlorobium tepidum were characterized by cloning and sequencing the genes which encode them, by overproducing the respective proteins in Escherichia coli, and by raising polyclonal antisera to the purified proteins. Three other proteins (AtpF, CT1970, and CT2144) which were identified in chlorosome fractions have similarly been characterized. The antisera were used to establish the distribution of each protein in various cellular fractions. Ten chlorosome proteins (CsmA, CsmB, CsmC, CsmD, CsmE, CsmF, CsmH, CsmI, CsmJ, and CsmX) copurified in a constant proportion together with bacteriochlorophyll c, and none of these 10 proteins was found in substantial amounts in other subcellular fractions. An antiserum to CsmH was highly effective in agglutinating chlorosomes, and antisera to CsmI, CsmJ, CsmX, and CsmA also immunoprecipitated chlorosomes to varying extents. However, an antiserum to CsmF did not agglutinate chlorosomes. The sequences of chlorosome proteins generally are not significantly similar to the sequences of other proteins in the databases. However, the N-terminal domains of three chlorosome proteins, CsmI, CsmJ, and CsmX, are related to adrenodoxin-type ferredoxins that ligate [2Fe-2S] clusters [Vassilieva, E. V., Antonkine, M. L., Zybailov, B. L., Yang, F., Jakobs, C. U., Golbeck, J. H., and Bryant, D. A. (2001) Biochemistry 40, 464-473]. The sequences of the C-terminal domains of these three proteins appear to be distantly related to CsmA and CsmE. The remaining chlorosome proteins can be divided into two additional structural families, CsmB/F and CsmC/D. CsmH is recovered in water-soluble form after overproduction in E. coli. Interestingly, this protein contains an N-terminal domain that is similar to CsmB/D, while its C-terminal domain is related to CsmC/D. The sequence relationships indicate that, although the protein composition of Chlorobium-type chlorosomes is superficially more complex than that of the chlorosomes of Chloroflexus aurantiacus, this heterogeneity is mostly produced by gene duplication and divergence among a small number of protein types.     

The phenylacetyl-CoA catabolon: a complex catabolic unit with broad biotechnological applications

The term catabolon was introduced to define a complex functional unit integrated by different catabolic pathways, which are, or could be, co-ordinately regulated, and that catalyses the transformation of structurally related compounds into a common catabolite. The phenylacetyl-CoA catabolon encompasses all the routes involved in the transformation of styrene, 2-phenylethylamine, trans-styrylacetic acid, phenylacetaldehyde, phenylacetic acid, phenylacetyl amides, phenylacetyl esters and n-phenylalkanoic acids containing an even number of carbon atoms, into phenylacetyl-CoA. This common intermediate is subsequently catabolized through a route of convergence, the phenylacetyl-CoA catabolon core, into general metabolites. The genetic organization of this central route, the biochemical significance of the whole functional unit and its broad biotechnological applications are discussed.     

From a short amino acidic sequence to the complete gene

A useful strategy directed to the isolation of a required gene with a high GC content is reported. Using a degenerate oligonucleotide probe, deduced from the amino terminus of a protein, it is possible to obtain a fragment of DNA containing its encoding gene by PCR amplification. Furthermore, the cloning of a desired gene can be accomplished in two steps by using an oligonucleotide deduced (i) from an internal sequence, (ii) from a consensus sequence, or (iii) from a DNA sequence adjacent to a disrupting element (transposon, insertion sequence, cassette). This method, which could be applied to a bacteriophage, plasmid, or cosmid genomic library, has been successfully used for cloning several genes from different biological systems.     

Ecological determinants of the occurrence and dynamics of Vibrio parahaemolyticus in offshore areas

The life cycle of Vibrio parahaemolyticus has been conventionally associated with estuarine areas characterized by moderate salinity and warm seawater temperatures. Recent evidence suggests that the distribution and population dynamics of V. parahaemolyticus may be shaped by the existence of an oceanic transport of communities of this organism mediated by zooplankton. To evaluate this possibility, the presence of V. parahaemolyticus in the water column of offshore areas of Galicia was investigated by PCR monthly over an 18-month period. Analysis of zooplankton and seawater showed that the occurrence of V. parahaemolyticus in offshore areas was almost exclusively associated with zooplankton and was present in 80% of the samples. The influence of environmental factors assessed by generalized additive models revealed that the abundance and seasonality of V. parahaemolyticus in zooplankton was favoured by the concurrence of downwelling periods that promoted the zooplankton patchiness. These results confirm that offshore waters may be common habitats for V. parahaemolyticus, including strains with virulent traits. Additionally, genetically related populations were found in offshore zooplankton and in estuaries dispersed along 1500 km. This finding suggests that zooplankton may operate as a vehicle for oceanic dispersal of V. parahaemolyticus populations, connecting distant regions and habitats, and thereby producing impacts on the local community demography and the spread of Vibrio-related diseases.     

Functional diversity across space and time: trends in wader communities on British estuaries

Aim British estuarine ecosystems support large populations of protected migratory waders. Understanding how wader communities vary spatially and how they may be changing temporally can greatly improve the understanding of these dynamic ecosystems. Here, we explore the variation in functional diversity (using a range of morphological and ecological traits) in order to identify the processes shaping wader communities on British estuaries and how these processes may be changing. Location England, Wales and Scotland. Methods We use national survey data (Wetland Bird Survey) from 1980/1981 to 2006/2007 winter to calculate functional diversity (FD) – an index that measures trait dispersion – in wader communities on 100 estuaries. We test for evidence of non‐random patterns of diversity and explore the relative importance of two key processes, environmental filtering and competition, in shaping these communities. Results The observed FD was significantly and positively associated with species richness and to a lesser extent estuary area, followed by longitude. An increase in observed FD was observed since 1980, supported by a small but significant slope. In the majority of cases, changes in FD were mirrored by changes in species richness. Observed FD was on average lower than expected by chance, as indicated by a negative value of observed minus expected FD. However, this difference became less negative over time, with observed minus expected FD values increasing slightly, but significantly, over the study period. Main conclusions Wader FD varies across British estuaries, and the relative influence of the processes by which communities are structured appears to be changing through time. We discuss the potential drivers underlying these patterns and the importance of identifying such drivers for the protection of wader communities.