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At least 451 records · Page 25

Transcriptome and Proteome Dynamics of a Light-Dark Synchronized Bacterial Cell Cycle

Growth of the ocean’s most abundant primary producer, the cyanobacterium Prochlorococcus, is tightly synchronized to the natural 24-hour light-dark cycle. We sought to quantify the relationship between transcriptome and proteome dynamics that underlie this obligate photoautotroph’s highly choreographed response to the daily oscillation in energy supply. Using RNA-sequencing transcriptomics and mass spectrometry-based quantitative proteomics, we measured timecourses of paired mRNA-protein abundances for 312 genes every 2 hours over a light-dark cycle. These temporal expression patterns reveal strong oscillations in transcript abundance that are broadly damped at the protein level, with mRNA levels varying on average 2.3 times more than the corresponding protein. The single strongest observed protein-level oscillation is in a ribonucleotide reductase, which may reflect a defense strategy against phage infection. The peak in abundance of most proteins also lags that of their transcript by 2–8 hours, and the two are completely antiphase for some genes. While abundant antisense RNA was detected, it apparently does not account for the observed divergences between expression levels. The redirection of flux through central carbon metabolism from daytime carbon fixation to nighttime respiration is associated with quite small changes in relative enzyme abundances. Our results indicate that expression responses to periodic stimuli that are common in natural ecosystems (such as the diel cycle) can diverge significantly between the mRNA and protein levels. Protein expression patterns that are distinct from those of cognate mRNA have implications for the interpretation of transcriptome and metatranscriptome data in terms of cellular metabolism and its biogeochemical impact.

54 ENVIRONMENTAL SCIENCES↗

The Bacterial Carbon-Fixing Organelle Is Formed by Shell Envelopment of Preassembled Cargo

Cyanobacteria play a significant role in the global carbon cycle. In Synechococcus elongatus, the carbon-fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is concentrated into polyhedral, proteinaceous compartments called carboxysomes. Using live cell fluorescence microscopy, we show that carboxysomes are first detected as small seeds of RuBisCO that colocalize with existing carboxysomes. These seeds contain little or no shell protein, but increase in RuBisCO content over several hours, during which time they are exposed to the solvent. The maturing seed is then enclosed by shell proteins, a rapid process that seals RuBisCO from the cytosol to establish a distinct, solvent-protected microenvironment that is oxidizing relative to the cytosol. These closure events can be spatially and temporally coincident with the appearance of a nascent daughter RuBisCO seed. Carboxysomes assemble in a stepwise fashion, inside-to-outside, revealing that cargo is the principle organizer of this compartment’s biogenesis. Our observations of the spatial relationship of seeds to previously formed carboxysomes lead us to propose a model for carboxysome replication via sequential fission, polymerization, and encapsulation of their internal cargo.

59 BASIC BIOLOGICAL SCIENCES↗

Proteomic Analysis of Cellular Response Induced by Multi-Walled Carbon Nanotubes Exposure in A549 Cells

The wide application of multi-walled carbon nanotubes (MWCNT) has raised serious concerns about their safety on human health and the environment. However, the potential harmful effects of MWCNT remain unclear and contradictory. To clarify the potentially toxic effects of MWCNT and to elucidate the associated underlying mechanisms, the effects of MWCNT on human lung adenocarcinoma A549 cells were examined at both the cellular and the protein level. Cytotoxicity and genotoxicity were examined, followed by a proteomic analysis (2-DE coupled with LC-MS/MS) of the cellular response to MWCNT. Our results demonstrate that MWCNT induces cytotoxicity in A549 cells only at relatively high concentrations and longer exposure time. Within a relatively low dosage range (30 mg/ml) and short time period (24 h), MWCNT treatment does not induce significant cytotoxicity, cell cycle changes, apoptosis, or DNA damage. However, at these low doses and times, MWCNT treatment causes significant changes in protein expression. A total of 106 proteins show altered expression at various time points and dosages, and of these, 52 proteins were further identified by MS. Identified proteins are involved in several cellular processes including proliferation, stress, and cellular skeleton organization. In particular, MWCNT treatment causes increases in actin expression. This increase has the potential to contribute to increased migration capacity and may be mediated by reactive oxygen species (ROS).

59 BASIC BIOLOGICAL SCIENCES↗

Designing Biological Systems for Sustainability and Programmed Environmental Interface (2011 JGI User Meeting)

The U.S. Department of Energy Joint Genome Institute (JGI) invited scientists interested in the application of genomics to bioenergy and environmental issues, as well as all current and prospective users and collaborators, to attend the annual DOE JGI Genomics of Energy & Environment Meeting held March 22-24, 2011 in Walnut Creek, Calif. The emphasis of this meeting was on the genomics of renewable energy strategies, carbon cycling, environmental gene discovery, and engineering of fuel-producing organisms. The meeting features presentations by leading scientists advancing these topics. Pam Silver of Harvard University gives a presentation on "Designing Biological Systems for Sustainability and Programmed Environmental Interface" at the 6th annual Genomics of Energy & Environment Meeting on March 23, 2011.

59 BASIC BIOLOGICAL SCIENCES↗

Space Station Freedom solar array containment box mechanisms

Space Station Freedom will feature six large solar arrays, called solar array wings, built by Lockheed Missiles & Space Company under contract to Rockwell International, Rocketdyne Division. Solar cells are mounted on flexible substrate panels which are hinged together to form a 'blanket.' Each wing is comprised of two blankets supported by a central mast, producing approximately 32 kW of power at beginning-of-life. During launch, the blankets are fan-folded and compressed to 1.5 percent of their deployed length into containment boxes. This paper describes the main containment box mechanisms designed to protect, deploy, and retract the solar array blankets: the latch, blanket restraint, tension, and guidewire mechanisms.

Johnson, Mark E.↗

Decorin modulates matrix mineralization in vitro

Decorin (DCN), a member of small leucine-rich proteoglycans, is known to modulate collagen fibrillogenesis. In order to investigate the potential roles of DCN in collagen matrix mineralization, several stable osteoblastic cell clones expressing higher (sense-DCN, S-DCN) and lower (antisense-DCN, As-DCN) levels of DCN were generated and the mineralized nodules formed by these clones were characterized. In comparison with control cells, the onset of mineralization by S-DCN clones was significantly delayed; whereas it was markedly accelerated and the number of mineralized nodules was significantly increased in As-DCN clones. The timing of mineralization was inversely correlated with the level of DCN synthesis. In these clones, the patterns of cell proliferation and differentiation appeared unaffected. These results suggest that DCN may act as an inhibitor of collagen matrix mineralization, thus modulating the timing of matrix mineralization.

NASA Discipline Musculoskeletal↗

Galacturonomannan and Golgi-derived membrane linked to growth and shaping of biogenic calcite

The coccolithophores are valuable models for the design and synthesis of composite materials, because the cellular machinery controlling the nucleation, growth, and patterning of their calcitic scales (coccoliths) can be examined genetically. The coccoliths are formed within the Golgi complex and are the major CaCO(3) component in limestone sediments-particularly those of the Cretaceous period. In this study, we describe mutants lacking a sulfated galacturonomannan and show that this polysaccharide in conjunction with the Golgi-derived membrane is directly linked to the growth and shaping of coccolith calcite but not to the initial orientated nucleation of the mineral phase.

NASA Discipline Cell Biology↗

Harold Kirby's symbionts of termites: karyomastigont reproduction and calonymphid taxonomy

Harold Kirby's brilliant principle of mastigont multiplicity is published here posthumously more than 40 years after it was written. He applies this principle to large multinucleate protist symbionts of termites in establishing the taxonomy of Calonymphids (Family Calonymphidae in Phylum Zoomastigina, Kingdom Protoctista). The nuclei and kinetosomes in these heterotrophic cells are organized into trichomonad-style mastigont units which reproduce independently of cytokinesis to generate nine new Calonympha and nineteen new Stephanonympha species. The total of six genera (Calonympha, Coronympha, Diplonympha, Metacoronympha, Snyderella and Stephanonympha, all symbionts of dry-wood-eating termites, Kalotermitidae) are recognized. With the aid of Michael Yamin, the distribution of all twenty-eight of Kirby's Calonympha and Stephanonympha species are tabulated. In italic type I have annotated this paper to be comprehensible to a wide readership of cell biologists, protistologists and those interested in insect symbionts. Although this extremely original and careful work was not finished when Kirby died suddenly in 1952, I deemed it important and complete enough to finally publish it so that it would not be lost to scientific posterity.

Non-NASA Center↗