Structural Analysis of Substrate, Reaction Intermediate, and Product Binding in Haemophilus influenzae Biotin Carboxylase

© 2015 American Chemical Society. Acetyl-CoA carboxylase catalyzes the first and regulated step in fatty acid synthesis. In most Gram-negative and Gram-positive bacteria, the enzyme is composed of three proteins: biotin carboxylase, a biotin carboxyl carrier protein (BCCP), and carboxyltransferase. The reaction mechanism involves two half-reactions with biotin carboxylase catalyzing the ATP-dependent carboxylation of biotin-BCCP in the first reaction. In the second reaction, carboxyltransferase catalyzes the transfer of the carboxyl group from biotin-BCCP to acetyl-CoA to form malonyl-CoA. In this report, high-resolution crystal structures of biotin carboxylase from Haemophilus influenzae were determined with bicarbonate, the ATP analogue AMPPCP; the carboxyphosphate intermediate analogues, phosphonoacetamide and phosphonoformate; the products ADP and phosphate; and the carboxybiotin analogue N1′-methoxycarbonyl biotin methyl ester. The structures have a common theme in that bicarbonate, phosphate, and the methyl ester of the carboxyl group of N1′-methoxycarbonyl biotin methyl ester all bound in the same pocket in the active site of biotin carboxylase and as such utilize the same set of amino acids for binding. This finding suggests a catalytic mechanism for biotin carboxylase in which the binding pocket that binds tetrahedral phosphate also accommodates and stabilizes a tetrahedral dianionic transition state resulting from direct transfer of CO2 from the carboxyphosphate intermediate to biotin

Genetic variation in male sexual behaviour in a population of white-footed mice in relation to photoperiod

In natural populations, genetic variation in seasonal male sexual behaviour could affect behavioural ecology and evolution. In a wild-source population of white-footed mice, Peromyscus leucopus, from Virginia, U.S.A., males experiencing short photoperiod show high levels of genetic variation in reproductive organ mass and neuroendocrine traits related to fertility. We tested whether males from two divergent selection lines, one that strongly suppresses fertility under short photoperiod (responder) and one that weakly suppresses fertility under short photoperiod (nonresponder), also differ in photoperiod-dependent sexual behaviour and responses to female olfactory cues. Under short, but not long, photoperiod, there were significant differences between responder and nonresponder males in sexual behaviour and likelihood of inseminating a female. Males that were severely oligospermic or azoospermic under short photoperiod failed to display sexual behaviour in response to an ovariectomized and hormonally primed receptive female. However, on the day following testing, females were positive for spermatozoa only when paired with a male having a sperm count in the normal range for males under long photoperiod. Males from the nonresponder line showed accelerated reproductive development under short photoperiod in response to urine-soiled bedding from females, but males from the responder line did not. The results indicate genetic variation in sexual behaviour that is expressed under short, but not long, photoperiod, and indicate a potential link between heritable neuroendocrine variation and male sexual behaviour. In winter in a natural population, this heritable behavioural variation could affect fitness, seasonal life history trade-offs and population growth. (C) 2015 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved

Age-Related Decline in Reproductive Sensitivity to Inhibition by Short Photoperiod in Peromyscus Leucopus

Seasonal environments favor the timing, of reproduction to match seasons when Successful reproduction is most likely. Most species of temperate zone mammals suppress reproduction in winter using changes in day length as a cue. In many species, individuals vary genetically in how strongly they respond to these seasonal cues. Individuals also may modify their response to day length depending upon other factors, including their age. Age-specific changes might occur because young, peripubertal rodents are more strongly affected by harsh conditions than adults, and therefore might be more sensitive to inhibitory photoperiods. We tested the hypothesis that genetic variation in responses to photoperiod persists as individuals age. Young males from a captive population of white-footed mice (Peromyscus leucopus) that is genetically variable for reproductive inhibition by short day length (SD) were tested for photoperiod responses. Mice were placed in SD within 3 days after birth, tested at age 70 days, allowed to mature for at least 18 weeks at long day length, and then tested again as adults aged \u3e= 34 weeks. Young males were more likely to be strongly reproductively Suppressed by SD than adults, indicating that age-specific changes in reproductive strategy occur in this Population. However, males that were reproductively photoresponsive when young also were more likely to be reproductively photoresponsive as adults. Thus, genetic tendency for reproductive sensitivity to photoperiod is a trait retained from puberty to adulthood, but attenuates with age

The three-dimensional structure of the biotin carboxylase-biotin carboxyl carrier protein complex of E. coli acetyl-CoA carboxylase

Acetyl-coenzyme A (acetyl-CoA) carboxylase is a biotin-dependent, multifunctional enzyme that catalyzes the regulated step in fatty acid synthesis. The Escherichia coli enzyme is composed of a homodimeric biotin carboxylase (BC), biotinylated biotin carboxyl carrier protein (BCCP), and an α2β2 heterotetrameric carboxyltransferase. This enzyme complex catalyzes two half-reactions to form malonyl-coenzyme A. BC and BCCP participate in the first half-reaction, whereas carboxyltransferase and BCCP are involved in the second. Three-dimensional structures have been reported for the individual subunits; however, the structural basis for how BCCP reacts with the carboxylase or transferase is unknown. Therefore, we report here the crystal structure of E. coli BCCP complexed with BC to a resolution of 2.49 Å. The protein-protein complex shows a unique quaternary structure and two distinct interfaces for each BCCP monomer. These BCCP binding sites are unique compared to phylogenetically related biotin-dependent carboxylases and therefore provide novel targets for developing antibiotics against bacterial acetyl-CoA carboxylase. © 2013 Elsevier Ltd

Integrating interprofessional electronic medical record teaching in preregistration healthcare degrees : A case study

Background Electronic medical record (EMR) adoption across healthcare necessitates a purposeful curriculum design to prepare graduates for the delivery of safe and effective patient care in digitally-enabled environments. Objective To describe the design and development of an Interprofessional Electronic Medical Record (iEMR) subject that introduces healthcare students to its utility in clinical settings. Methods A six-stage design-based educational research framework (Focus, Formulation, Contextualisation, Definition, Implementation, Evaluation) was used to instigate the iEMR design and development in nursing and five allied health graduate entry to practice (preregistration) degrees at an Australian university. Results In the Focus process, the concept and interdisciplinary partnerships were developed. The Formulation process secured grant support for subject design and development, including a rapid literature review to accommodate various course and curriculum structures. Discipline-specific subject themes were created through the Contextualisation process. During the Definition process, learning objectives and content resources were built. The Implementation process describes the pilot implementation in the nursing program, where assessment tasks were refined, and interdisciplinary clinical case studies originated. Discussion The design and development of an iEMR subject is underpinned by internal support for educational innovation and in alignment with digital health strategies in employer organisations. Identified barriers include faculty-level changes in strategic support for teaching innovation, managerial expectations of workload, the scope of work required by academics and learning designers, and the gap between the technology platform required to support online learning and the infrastructure needed to support simulated EMR use. A key discovery was the difficulty of finding EMR software, whether designed for teaching purposes or for clinical use, that could be adapted to meet the needs of this project. Conclusion The lessons learned are relevant to educators and learning designers attempting a similar process. Issues remain surrounding the sustainability of the iEMR subject and maintaining academic responsibility for ongoing curriculum management

Cluster M Mycobacteriophages Bongo, PegLeg, and Rey with Unusually Large Repertoires of tRNA Isotopes

Genomic analysis of a large set of phages infecting the common hostMycobacterium smegmatis mc2155 shows that they span considerable genetic diversity. There are more than 20 distinct types that lack nucleotide similarity with each other, and there is considerable diversity within most of the groups. Three newly isolated temperate mycobacteriophages, Bongo, PegLeg, and Rey, constitute a new group (cluster M), with the closely related phages Bongo and PegLeg forming subcluster M1 and the more distantly related Rey forming subcluster M2. The cluster M mycobacteriophages have siphoviral morphologies with unusually long tails, are homoimmune, and have larger than average genomes (80.2 to 83.7 kbp). They exhibit a variety of features not previously described in other mycobacteriophages, including noncanonical genome architectures and several unusual sets of conserved repeated sequences suggesting novel regulatory systems for both transcription and translation. In addition to containing transfer-messenger RNA and RtcB-like RNA ligase genes, their genomes encode 21 to 24 tRNA genes encompassing complete or nearly complete sets of isotypes. We predict that these tRNAs are used in late lytic growth, likely compensating for the degradation or inadequacy of host tRNAs. They may represent a complete set of tRNAs necessary for late lytic growth, especially when taken together with the apparent lack of codons in the same late genes that correspond to tRNAs that the genomes of the phages do not obviously encode

Precision pulse shape simulation for proton detection at the Nab experiment

The Nab experiment at Oak Ridge National Laboratory, USA, aims to measure the beta-antineutrino angular correlation following neutron $\beta$ decay to an anticipated precision of approximately 0.1\%. The proton momentum is reconstructed through proton time-of-flight measurements, and potential systematic biases in the timing reconstruction due to detector effects must be controlled at the nanosecond level. We present a thorough and detailed semiconductor and quasiparticle transport simulation effort to provide precise pulse shapes, and report on relevant systematic effects and potential measurement schemes

US Cosmic Visions: New Ideas in Dark Matter 2017: Community Report

This white paper summarizes the workshop "U.S. Cosmic Visions: New Ideas in Dark Matter" held at University of Maryland on March 23-25, 2017.Comment: 102 pages + reference