Distribution of the catabolic transposon Tn5271 in a groundwater bioremediation system.

The distribution of Tn5271-related DNA sequences in samples of groundwater and a groundwater bioremediation system at the Hyde Park (Niagara Falls, N.Y.) chemical landfill site was investigated. PCR amplification of target sequences within the cha genes of Tn5271 revealed similar sequences in the groundwater community and in samples from the sequencing batch reactors treating that groundwater. Cell dilution combined with PCR amplification indicated that cha sequences were carried in about 1 of 10 culturable bacteria from the treatment system. Characterization of isolates involved in chlorobenzoate and toluene biodegradation in the treatment system indicated that two phenotypic clusters, Alcaligenes faecalis type 2 and CDC group IVC-2, contained all of the Tn5271 probe-positive isolates from the community. These two groups differed phenotypically from recipient groups isolated following horizontal transfer of pBRC60 (Tn5271) in pristine freshwater microcosms. A genetic rearrangement in Tn5271 attributable to the intramolecular transposition of the flanking element IS1071R was detected in an isolate from the treatment system. Comparison of the structure of the intramolecular transposition derivative from groundwater isolate OCC13(pBRC13) with a laboratory-derived intramolecular transposition derivative of pBRC60 revealed similarities. The rearrangement was shown to increase the stability of the plasmid under starvation conditions.     

Diversity and conservation of blackwater fishes in Peninsular Malaysia,particularly in the North Selangor peat swamp forest

One of the most extreme freshwater habitats in Peninsular Malaysia is the peat swamp forest, with dark-coloured and highly acidic waters. Surprisingly, little is known about blackwater fishes in Peninsular Malaysia. Until 1968, only 26 fish species were known from blackwaters throughout Peninsular Malaysia, of which only one can be regarded as stenotopic. A recent intensive survey of part of the North Selangor peat swamp forest yielded 47 species, of which 14 are probably stenotopic taxa. These include four undescribed species and several new records for western Peninsular Malaysia. These discoveries are significant in that they include the family Chaudhuriidae which until 1985, was not reported from Sundaic Southeast Asia, and the rare genus Encheloclarias which had not been encountered for over 50 years. The rapid rate of destruction of the peat swamp forest owing to development, forestry and agricultural activities must be halted or slowed significantly to enable the proper zoological surveys and studies to be conducted. Conservation plans and environmental impact assessments based on inadequate sampling and knowledge of species present is acutely dangerous. There are no longer substantial undisturbed blackwater peat swamp forests left in most of Peninsular Malaysia. Conservation of the remaining blackwater biotopes is critically important if extinction of many species, here regarded as economically valuable renewable resources, is to be prevented.     

The systematics and ecology of snakeheads (Pisces: Channidae) in Peninsular Malaysia and Singapore

Seven species of snakeheads (Channidae) are known from Peninsular Malaysia and Singapore, viz. Channa bankanensis, C. gachua, C. lucius, C. marulioides, C. melasoma, C. micropeltes and C. striata. Up-to-date distribution maps of each species are presented, including new records. Their systematics is reviewed and partially revised. The taxonomic status of C. marulioides and C. melanoptera is clarified. Specimens from Peninsular Malaysia identified as C. melanoptera sensu Weber & de Beaufort, 1922, proved to be the adult form of C. marulioides s.str. The real C. melanoptera appears to be restricted to Borneo and possibly Sumatra. The life history of a blackwater species C. bankanensis is also documented, with regards to the morphological and colour-pattern changes associated with growth. An updated key to the seven species based on morphometric measurements and meristic counts is presented.     

Phosphorylation and Palmitoylation of the Human D2L Dopamine Receptor in Sf9 Cells

Abstract: We have expressed and biochemically characterized the human D2_long (D2_L) dopamine receptor isoform using the baculovirus/Sf9 cell system. The expressed receptor bound ligands with a pharmacological profile similar to that reported for neuronal and cloned D2_L receptors expressed in mammalian cell lines. Dopamine binding to D2_L receptor was sensitive to guanine nucleotides, indicating receptor coupling to endogenous G proteins. A D2_L receptor-specific antibody identified two major protein species at ∼44 kDa and at ∼93 kDa in immunoblots, suggesting the presence of D2_L receptor monomers and dimers. Both species were purified by immunoprecipitation from digitonin-solubilized preparation of cells expressing D2_L receptor prelabeled with ³²P_i or [³H]-palmitate. These results constitute the first direct evidence for D2_L receptor phosphorylation and palmitoylation.     

The α2(XI) collagen gene lies within 8 kb of Pb in the proximal portion of the murine major histocompatibility complex

A number of serious hereditary disorders are now known to be associated with defective expression of collagen genes, and these findings have underscored the important and varied roles that the collagen family of genes must play during normal mammalian development. Although the activities of genes encoding the quantitatively major types of collagen are fairly well characterized, functions of the many minor types of collagen remain a matter of speculation. As a first step toward a functional analysis of type XI collagen, a member of this class of poorly understand minor collagen proteins which is expressed primarily in hyaline cartilage, we have used human probes for the gene encoding the protein's 2-subunit (COL11A2) to isolate and map homologous murine DNA sequences. Our results demonstrate that Col11a-2 is embedded within the major histocompatibility complex (MHC), within 8.4 kb of the class II pseudogene locus, Pb, and confirm that human and murine 2(XI) collagen genes are located in very similar genomic environments. The conserved location of these genes raises the possibility that type XI collagen genes may contribute to one or more of the diverse hereditary disorders known to be linked to the MHC in mouse and human.     

Ras transformation results in an elevated level of cyclin D1 and acceleration of G1 progression in NIH 3T3 cells.

Ectopic overexpression of v-H-Ras protein in NIH 3T3 cells resulted in cellular transformation and an acceleration of G1 progression of these cells. A shortened G1 phase was found to be associated with an increased level of cyclin D1 but not cyclin E protein. Using an antisense blocking method, reduced synthesis of cyclin D1 in v-H-Ras transformants resulted in a slower G1 progression rate of these cells. Although constitutive overexpression of cyclin D1 in NIH 3T3 cells accelerated G1 progression, cells remained untransformed. Furthermore, inhibition of cyclin D1 synthesis greatly impaired the soft-agar cloning efficiency of v-H-Ras transformants. These results suggest that increased expression of cyclin D1 is necessary but not sufficient for the transforming activity of v-H-Ras. Similar effect on cell cycle progression was also observed in Raf-transformed cells. In addition to cyclin D1, cyclin E protein was found to be elevated in Src transformants. This may account for the further shortening of the G1 phase of these cells. Activation of an additional Ras-independent pathway was suggested to be responsible for the further acceleration of the G1 phase in Src transformants.