Experimental Genome-Wide Determination of RNA Polyadenylation in \u3cem\u3eChlamydomonas reinhardtii\u3c/em\u3e

The polyadenylation of RNA is a near-universal feature of RNA metabolism in eukaryotes. This process has been studied in the model alga Chlamydomonas reinhardtii using low-throughput (gene-by-gene) and high-throughput (transcriptome sequencing) approaches that recovered poly(A)-containing sequence tags which revealed interesting features of this critical process in Chlamydomonas. In this study, RNA polyadenylation has been studied using the so-called Poly(A) Tag Sequencing (PAT-Seq) approach. Specifically, PAT-Seq was used to study poly(A) site choice in cultures grown in four different media types—Tris-Phosphate (TP), Tris-Phosphate-Acetate (TAP), High-Salt (HS), and High-Salt-Acetate (HAS). The results indicate that: 1. As reported before, the motif UGUAA is the primary, and perhaps sole, cis-element that guides mRNA polyadenylation in the nucleus; 2. The scope of alternative polyadenylation events with the potential to change the coding sequences of mRNAs is limited; 3. Changes in poly(A) site choice in cultures grown in the different media types are very few in number and do not affect protein-coding potential; 4. Organellar polyadenylation is considerable and affects primarily ribosomal RNAs in the chloroplast and mitochondria; and 5. Organellar RNA polyadenylation is a dynamic process that is affected by the different media types used for cell growth

Methods Used to Evaluate Employment and Training Programs in the Past

The authors begin with a thorough assessment of the many nonexperimental employment and training program evaluation techniques based on non-random comparison groups. These techniques typically use econometric methods to estimate the effects of employment and training programs by using comparison groups from non-program external sources. Then, recognizing the inherent drawbacks in these methods, Bell, Orr, Blomquist and Cain respond by reintroducing an evaluation method first implemented in the 1960s, the use of internal comparison groups consisting of nonparticipating program applicants. These groups include withdrawals, screen-outs and no-shows of the programs being evaluated in order to solve the selection bias problem. By applying to the program, say the authors, nonparticipating applicants reveal themselves to have some of the same difficult-to-measure, personal characteristics that inspire participants to seek help in response to their current economic situation. The methodology of this technique is updated, then tested against the random experimental findings derived from a controlled job training experiment, the AFDC Homemaker-Home Health Aide Demonstrations. Encouraging results are presented along with useful suggestions for designers and implementers of all types of program evaluations.https://research.upjohn.org/up_press/1080/thumbnail.jp

The sub-cellular localization of Sulfolobus DNA replication

Analyses of the DNA replication-associated proteins of hyperthermophilic archaea have yielded considerable insight into the structure and biochemical function of these evolutionarily conserved factors. However, little is known about the regulation and progression of DNA replication in the context of archaeal cells. In the current work, we describe the generation of strains of Sulfolobus solfataricus and Sulfolobus acidocaldarius that allow the incorporation of nucleoside analogues during DNA replication. We employ this technology, in conjunction with immunolocalization analyses of replisomes, to investigate the sub-cellular localization of nascent DNA and replisomes. Our data reveal a peripheral localization of replisomes in the cell. Furthermore, while the two replication forks emerging from any one of the three replication origins in the Sulfolobus chromosome remain in close proximity, the three origin loci are separated

Leionema lamprophyllum subsp. fractum (Rutaceae); a new and highly restricted taxon from the Hunter Valley of New South Wales

A new subspecies of Leionema lamprophyllum, formerly included in L. lamprophyllum subsp. obovatum F.M.Anderson, is here described as new. Currently known from fewer than 50 individuals, the new subsp. is highly restricted and warrants a conservation risk code of at least Endangered. An illustration of the new taxon, notes on its distribution and habitat, and a key to all four subspecies of L. lamprophyllum are also provided

Demonstrating superior discrimination of locally prepared states using nonlocal measurements

We experimentally demonstrate the superior discrimination of separated, unentangled two-qubit correlated states using nonlocal measurements, when compared with measurements based on local operations and classical communications. When predicted theoretically, this phenomenon was dubbed quantum nonlocality without entanglement. We characterize the performance of the nonlocal, or joint, measurement with a payoff function, for which we measure 0.72 +/- 0.02, compared with the maximum locally achievable value of 2/3 and the overall optimal value of 0.75

Visualization of replication initiation and elongation in Drosophila

Chorion gene amplification in the ovaries of Drosophila melanogaster is a powerful system for the study of metazoan DNA replication in vivo. Using a combination of high-resolution confocal and deconvolution microscopy and quantitative realtime PCR, we found that initiation and elongation occur during separate developmental stages, thus permitting analysis of these two phases of replication in vivo. Bromodeoxyuridine, origin recognition complex, and the elongation factors minichromosome maintenance proteins (MCM)2–7 and proliferating cell nuclear antigen were precisely localized, and the DNA copy number along the third chromosome chorion amplicon was quantified during multiple developmental stages. These studies revealed that initiation takes place during stages 10B and 11 of egg chamber development, whereas only elongation of existing replication forks occurs during egg chamber stages 12 and 13. The ability to distinguish initiation from elongation makes this an outstanding model to decipher the roles of various replication factors during metazoan DNA replication. We utilized this system to demonstrate that the pre–replication complex component, double-parked protein/cell division cycle 10–dependent transcript 1, is not only necessary for proper MCM2–7 localization, but, unexpectedly, is present during elongation