The Rhodococcus opacus PD630 Heparin-Binding Hemagglutinin Homolog TadA Mediates Lipid Body Formation ▿

Generally, prokaryotes store carbon as polyhydroxyalkanoate, starch, or glycogen. The Gram-positive actinomycete Rhodococcus opacus strain PD630 is noteworthy in that it stores carbon in the form of triacylglycerol (TAG). Several studies have demonstrated that R. opacus PD630 can accumulate up to 76% of its cell dry weight as TAG when grown under nitrogen-limiting conditions. While this process is well studied, the underlying molecular and biochemical mechanisms leading to TAG biosynthesis and subsequent storage are poorly understood. We designed a high-throughput genetic screening to identify genes and their products required for TAG biosynthesis and storage in R. opacus PD630. We identified a gene predicted to encode a putative heparin-binding hemagglutinin homolog, which we have termed tadA (triacylglycerol accumulation deficient), as being important for TAG accumulation. Kinetic studies of TAG accumulation in both the wild-type (WT) and mutant strains demonstrated that the tadA mutant accumulates 30 to 40% less TAG than the parental strain (WT). We observed that lipid bodies formed by the mutant strain were of a different size and shape than those of the WT. Characterization of TadA demonstrated that the protein is capable of binding heparin and of agglutinating purified lipid bodies. Finally, we observed that the TadA protein localizes to lipid bodies in R. opacus PD630 both in vivo and in vitro. Based on these data, we hypothesize that the TadA protein acts to aggregate small lipid bodies, found in cells during early stages of lipid storage, into larger lipid bodies and thus plays a key role in lipid body maturation in R. opacus PD630

Morphological and molecular characterization of common bean landraces and cultivars from the Caribbean

Little information is available regarding the relationship of Caribbean bean landraces with the Andean and Mesoamerican gene pools of common bean (Phaseolus vulgaris L.) While small-seeded Mesoamerican black beans are prevalent in parts of the Caribbean, many of the regionally preferred red mottled and medium-to-large-seeded bean landraces found there are postulated to belong to the Andean gene pool. To test this hypothesis, we analyzed the morphological characteristics, phenological traits, phaseolin status, and random amplified polymorphic DNA (RAPD) banding patterns of 54 red mottled or large-seeded bean landraces and cultivars from the Caribbean (16 from the Dominican Republic, 14 from Haiti, 1 from Jamaica and 23 from Puerto Rico) compared with 11 Andean bean lines from other regions. To estimate phylogenetic relationships among the lines, distances were calculated and dendrograms constructed for morphological and/or phenological and molecular characteristics by the complete clustering method. The landraces were grouped into two clusters morphologically: one with Mesoamerican characteristics, which included all the red mottled lines from Haiti and three landraces from the Dominican Republic collected near the Haitian border and the other with Andean characteristics, which included all the lines from Puerto Rico and the remaining lines from the Dominican Republic. RAPD and phaseolin polymorphisms identified three groups, one corresponding to the genotypes with Mesoamerican morphologies, one to those with Andean morphologies, and another that had Andean phenotypes but proximity to the Mesoamerican group, suggesting possible introgression between the gene pools. Phaseolin variability agreed with other molecular and morphological data allowing the assignment of genotypes to a Mesoamerican or Andean gene pool. Knowledge of this variability may provide useful information concerning the potential value of Caribbean landraces to Andean bean breeding programs

Longitudinal Impact of WTC Dust Inhalation on Rat Cardiac Tissue Transcriptomic Profiles

First responders (FR) exposed to the World Trade Center (WTC) Ground Zero air over the first week after the 9/11 disaster have an increased heart disease incidence compared to unexposed FR and the general population. To test if WTC dusts were causative agents, rats were exposed to WTC dusts (under isoflurane [ISO] anesthesia) 2 h/day on 2 consecutive days; controls received air/ISO or air only. Hearts were collected 1, 30, 240, and 360 d post-exposure, left ventricle total RNA was extracted, and transcription profiles were obtained. The data showed that differentially expressed genes (DEG) for WTC vs. ISO rats did not reach any significance with a false discovery rate (FDR) < 0.05 at days 1, 30, and 240, indicating that the dusts did not impart effects beyond any from ISO. However, at day 360, 14 DEG with a low FDR were identified, reflecting potential long-term effects from WTC dust alone, and the majority of these DEG have been implicated as having an impact on heart functions. Furthermore, the functional gene set enrichment analysis (GSEA) data at day 360 showed that WTC dust could potentially impact the myocardial energy metabolism via PPAR signaling and heart valve development. This is the first study showing that WTC dust could significantly affect some genes that are associated with the heart/CV system, in the long term. Even > 20 years after the 9/11 disaster, this has potentially important implications for those FR exposed repeatedly at Ground Zero over the first week after the buildings collapsed