Strong Sequence Research Articles

A role for the Drosophila zinc transporter Zip88E in protecting against dietary zinc toxicity.

Zinc absorption in animals is thought to be regulated in a local, cell autonomous manner with intestinal cells responding to dietary zinc content. The Drosophila zinc transporter Zip88E shows strong sequence similarity to Zips 42C.1, 42C.2 and 89B as well as mammalian Zips 1, 2 and 3, suggesting that it may act in concert with the apically-localised Drosophila zinc uptake transporters to facilitate dietary zinc absorption by importing ions into the midgut enterocytes. However, the functional characterisation of Zip88E presented here indicates that Zip88E may instead play a role in detecting and responding to zinc toxicity. Larvae homozygous for a null Zip88E allele are viable yet display heightened sensitivity to elevated levels of dietary zinc. This decreased zinc tolerance is accompanied by an overall decrease in Metallothionein B transcription throughout the larval midgut. A Zip88E reporter gene is expressed only in the salivary glands, a handful of enteroendocrine cells at the boundary between the anterior and middle midgut regions, and in two parallel strips of sensory cell projections connecting to the larval ventral ganglion. Zip88E expression solely in this restricted subset of cells is sufficient to rescue the Zip88E mutant phenotype. Together, our data suggest that Zip88E may be functioning in a small subset of cells to detect excessive zinc levels and induce a systemic response to reduce dietary zinc absorption and hence protect against toxicity.

A taxonomic revision of the Cardiocondyla nuda group (Hymenoptera: Formicidae)

A taxonomic revision of the Cardiocondyla nuda species group is presented based on methods of Numeric Morphology-Based Alpha-Taxonomy (NUMOBAT) and supplemented by analysis of mtDNA. A total of 258 samples with 571 worker individuals were investigated by the hierarchical and non-hierarchical exploratory data analyses NC-Ward and NC-K-Means clustering considering 16 NUMOBAT characters. Two species are described as new, increasing the number of species in the group to eight. We separate the group into two main clades: the C. mauritanica species complex, which is of Oriental and Indo-Australian origin and contains the cryptic species C. mauritanica Forel 1890, C. strigifrons Viehmeyer 1922, C. kagutsuchi Terayama 1999, and C. itsukii sp. nov. and the Australasian and Polynesian C. nuda species complex with the cryptic species C. nuda (Mayr 1866), C. atalanta Forel 1915, C. paranuda Seifert 2003, and C. compressa sp. nov. The mean error of the two NC-clustering methods relative to the controlling linear discriminant analysis was 0.4% in C. mauritanica, 2.2% in C. itsukii, 0% in C. strigifrons, 0% in C. kagutsuchi, 1.5% in C. nuda, 3.2% in C. atalanta and 3.2% in C. paranuda—all these data are below the 4% threshold recommended by the Pragmatic Species Concept. The morphologically determined species clusters were confirmed by mtDNA data with a rather strong sequence divergence among the cryptic species of the C. nuda complex of 5.6–7.9%. The mean mismatch of two different mtDNA analyses with NUMOBAT clustering was 5.4% in 54 samples of seven species of the C. nuda group for which mtDNA data were available. The mismatch thus is smaller than in many other studies of Eumetazoa in general or ants in particular and is probably explained by low frequencies of ancient hybridization and/or incomplete lineage sorting. Comments on zoogeography, colony demography and behavior are given in the species sections and determination keys are provided. Cardiocondyla ectopia Snelling 1974 and Leptothorax caparica Henin, Paiva & Collingwood 2002 (syn nov.) are synonymized under Cardiocondyla mauritanica. Cardiocondyla nuda sculptinodis is not a member of the C. nuda group and is moved to C. shuckardi sculptinodis, a revived combination.

Structure, function and evolution of the cyanobacterial orange carotenoid protein and its homologs.

Contents 937 I. 937 II. 938 III. 939 IV. 943 V. 947 VI. 948 948 References 949 SUMMARY: The orange carotenoid protein (OCP) is a water-soluble, photoactive protein involved in thermal dissipation of excess energy absorbed by the light-harvesting phycobilisomes (PBS) in cyanobacteria. The OCP is structurally and functionally modular, consisting of a sensor domain, an effector domain and a keto-carotenoid. On photoactivation, the OCP converts from a stable orange form, OCPO , to a red form, OCPR . Activation is accompanied by a translocation of the carotenoid deeper into the effector domain. The increasing availability of cyanobacterial genomes has enabled the identification of new OCP families (OCP1, OCP2, OCPX). The fluorescence recovery protein (FRP) detaches OCP1 from the PBS core, accelerating its back-conversion to OCPO ; by contrast, other OCP families are not regulated by FRP. N-terminal domain homologs, the helical carotenoid proteins (HCPs), have been found among diverse cyanobacteria, occurring as multiple paralogous groups, with two representatives exhibiting strong singlet oxygen (1 O2 ) quenching (HCP2, HCP3) and another capable of dissipating PBS excitation (HCP4). Crystal structures are presently available for OCP1 and HCP1, and models of other HCP subtypes can be readily produced as a result of strong sequence conservation, providing new insights into the determinants of carotenoid binding and 1 O2 quenching.

Sequence signal processing of 2012 Ahar–Varzaghan earthquake (M W 6.4) of NW Iran

Before starting seismic cycle of Ahar–Varzaghan 2012 event, a partial gap in the form of a pre-seismic calm sequence (seismicity rate, r = 0.46 event/year, b = 1.4) with duration of 303 days spatially has dominated over the entire seismogenic area. From April 17, 2012, to May 31, 2012, r significantly increased to 2.16, indicating strong foreshock sequence, and b value changed to 1.9, remarkably. In the last two months before the mainshock, foreshocks have partially migrated toward the earthquake fault (with a decrease in size, b = 2.0). Significantly, high rate of seismicity and low V P /V S (1.64) in the foreshocks sequence and also very high seismicity rate (17.3) and high V P /V S (1.76) in the aftershocks sequence make substantial differences between the seismic cycle and the background seismicity. Moreover, a significant E–W migration of the microseismicity was confirmed in the study area.

Identification of transcripts with short stuORFs as targets for DENR\u2022MCTS1-dependent translation in human cells

The non-canonical initiation factors DENR and MCTS1 have been linked to cancer and autism. We recently showed in Drosophila that DENR and MCTS1 regulate translation re-initiation on transcripts containing upstream Open Reading Frames (uORFs) with strong Kozak sequences (stuORFs). Due to the medical relevance of DENR and MCTS1, it is worthwhile identifying the transcripts in human cells that depend on DENR and MCTS1 for their translation. We show here that in humans, as in Drosophila, transcripts with short stuORFs require DENR and MCTS1 for their optimal expression. In contrast to Drosophila, however, the dependence on stuORF length in human cells is very strong, so that only transcripts with very short stuORFs coding for 1 amino acid are dependent on DENR and MCTS1. This identifies circa 100 genes as putative DENR and MCTS1 translational targets. These genes are enriched for neuronal genes and G protein-coupled receptors. The identification of DENR and MCTS1 target transcripts will serve as a basis for future studies aimed at understanding the mechanistic involvement of DENR and MCTS1 in cancer and autism.

Biosynthesis of 1α-hydroxycorticosterone in the winter skate Leucoraja ocellata: evidence to suggest a novel steroidogenic route.

The present study explores the ability of intracellular bacteria within the renal-inter-renal tissue of the winter skate Leucoraja ocellata to metabolize steroids and contribute to the synthesis of the novel elasmobranch corticosteroid, 1α-hydroxycorticosterone (1α-OH-B). Despite the rarity of C1 hydroxylation noted in the original identification of 1α-OH-B, literature provides evidence for steroid C1 hydroxylation by micro-organisms. Eight ureolytic bacterial isolates were identified in the renal-inter-renal tissue of L. ocellata, the latter being the site of 1α-OH-B synthesis. From incubations of bacterial isolates with known amounts of potential 1α-OH-B precursors, one isolate UM008 of the genus Rhodococcus was seen to metabolize corticosteroids and produce novel products via HPLC analysis. Cations Zn2+ and Fe3+ altered metabolism of certain steroid precursors, suggesting inhibition of Rhodococcus steroid catabolism. Genome sequencing of UM008 identified strong sequence and structural homology to that of Rhodococcus erythropolis PR4. A complete enzymatic pathway for steroid-ring oxidation as documented within other Actinobacteria was identified within the UM008 genome. This study highlights the potential role of Rhodococcus bacteria in steroid metabolism and proposes a novel alternative pathway for 1α-OH-B synthesis, suggesting a unique form of mutualism between intracellular bacteria and their elasmobranch host.

Numerical Investigation of Four Seismic Upgrading Techniques on a Masonry Bell Tower

This paper presents some preliminary results from the numerical investigations of different interventions for the seismic upgrading of a masonry bell tower. Masonry bell towers belong to the cultural and historical heritage, are very widespread in Italy and their conservation has been given special attention in recent years. Their high vulnerability is related to the combination of mechanical material properties, geometrical irregularities, presence of inclination, foundation settlements and seismicity of the area. The present tower is located in Emilia Romagna Region, Northern Italy, recently stricken by a strong seismic sequence in May 2012. Analyses results show that the retrofitting of masonry bell towers is highly recommended in order to mitigate their present seismic risk. Four different upgrading techniques, consisting of grouted mortar, FRP sheets, internal steel frame and prestressing vertical rods, are investigated and compared in terms of structural performance improvement. The grouted mortar and internal steel frame techniques are found to be reasonable solutions when compared to the other proposals for the specific seismic intensity level. Furthermore, the other techniques have a relatively high level of invasiveness which is recommended to be avoid where possible. Experience has shown that the most effective interventions are those that not only protect the structure, but also preserve the built heritage.

AnCo3, a New Member of the Emerging Family of Phage-Like Plasmids.

ABSTRACTA phage-like plasmid isolated from a clinical isolate of Salmonella enterica serovar Derby has strong nucleotide sequence identity to the phage-like plasmids pSTM_phi isolated from Salmonella enterica serovar Typhimurium L495, AnCo1 and AnCo2 from Escherichia coli 243 and Escherichia coli 244, and the virulent Salmonella-specific SSU5 bacteriophage.

Complete Genome Sequences of Two Phage-Like Plasmids Carrying the CTX-M-15 Extended-Spectrum β-Lactamase Gene.

ABSTRACTTwo similar phage-like plasmids carrying CTX-M-15 resistance cassettes were identified from two environmental Escherichia coli isolates. They demonstrate strong nucleotide sequence identity to the phage-like plasmid pECOH89 and Salmonella bacteriophage SSU5.

Human Immunoglobulin Heavy Gamma Chain Polymorphisms: Molecular Confirmation Of Proteomic Assessment

Immunoglobulin G (IgG) proteins are known for the huge diversity of the variable domains of their heavy and light chains, aimed at protecting each individual against foreign antigens. The IgG also harbor specific polymorphism concentrated in the CH2 and CH3-CHS constant regions located on the Fc fragment of their heavy chains. But this individual particularity relies only on a few amino acids among which some could make accurate sequence determination a challenge for mass spectrometry-based techniques.The purpose of the study was to bring a molecular validation of proteomic results by the sequencing of encoding DNA fragments. It was performed using ten individual samples (DNA and sera) selected on the basis of their Gm (gamma marker) allotype polymorphism in order to cover the main immunoglobulin heavy gamma (IGHG) gene diversity. Gm allotypes, reflecting part of this diversity, were determined by a serological method. On its side, the IGH locus comprises four functional IGHG genes totalizing 34 alleles and encoding the four IgG subclasses. The genomic study focused on the nucleotide polymorphism of the CH2 and CH3-CHS exons and of the intron. Despite strong sequence identity, four pairs of specific gene amplification primers could be designed. Additional primers were identified to perform the subsequent sequencing. The nucleotide sequences obtained were first assigned to a specific IGHG gene, and then IGHG alleles were deduced using a home-made decision tree reading of the nucleotide sequences. IGHG amino acid (AA) alleles were determined by mass spectrometry. Identical results were found at 95% between alleles identified by proteomics and those deduced from genomics. These results validate the proteomic approach which could be used for diagnostic purposes, namely for a mother-and-child differential IGHG detection in a context of suspicion of congenital infection.

A Specific Mutational Signature Associated with DNA 8-Oxoguanine Persistence in MUTYH-defective Colorectal Cancer

8-Oxoguanine, a common mutagenic DNA lesion, generates G:C>T:A transversions via mispairing with adenine during DNA replication. When operating normally, the MUTYH DNA glycosylase prevents 8-oxoguanine-related mutagenesis by excising the incorporated adenine. Biallelic MUTYH mutations impair this enzymatic function and are associated with colorectal cancer (CRC) in MUTYH-Associated Polyposis (MAP) syndrome. Here, we perform whole-exome sequencing that reveals a modest mutator phenotype in MAP CRCs compared to sporadic CRC stem cell lines or bulk tumours. The excess G:C>T:A transversion mutations in MAP CRCs exhibits a novel mutational signature, termed Signature 36, with a strong sequence dependence. The MUTYH mutational signature reflecting persistent 8-oxoG:A mismatches occurs frequently in the APC, KRAS, PIK3CA, FAT4, TP53, FAT1, AMER1, KDM6A, SMAD4 and SMAD2 genes that are associated with CRC. The occurrence of Signature 36 in other types of human cancer indicates that DNA 8-oxoguanine-related mutations might contribute to the development of cancer in other organs.

Abstract A46: Nup153 and Nup50 promote recruitment of 53BP1 to DNA repair foci by antagonizing BRCA1-dependent events

Abstract This study focuses on an unexpected role for particular nuclear pore proteins in the antagonism between the DNA repair factors BRCA1 and 53BP1, and the impact this has on the response to poly(A) ribose polymerase (PARP) inhibition. Repair of DNA double strand breaks, a cornerstone of genomic integrity, typically follows one of two distinct pathways: the high fidelity process of homologous recombination (HR) repair, in which BRCA1 plays a key role, and the more error-prone process of non-homologous end joining (NHEJ), which relies on 53BP1. The balance between NHEJ and HR depends, in part, on whether 53BP1 predominates in binding to damage sites, where it protects the DNA ends from resection and prevents HR from occurring. Conversely, BRCA1 binding counteracts 53BP1-mediated events to promote HR repair. While mutual antagonism between BRCA1 and 53BP1 has been appreciated for some time, factors that modulate this relationship are only beginning to be elucidated. The nuclear pore component Nup153 has previously been implicated in the DNA damage response, specifically in promoting nuclear import of 53BP1. To further explore the role of Nup153 in this context, we engineered an additional strong nuclear localization sequence (NLS) on 53BP1 and found that this overrides the requirement for Nup153 in nuclear import of 53BP1. While 53BP1-NLS is recruited normally to DNA damage foci in control-treated cells, its accumulation at these sites is still severely impaired in Nup153-depleted cells, indicating that Nup153 has an additional and distinct role in 53BP1 intranuclear targeting. These observations are further underscored by the additional finding that a second nuclear pore protein, Nup50, is also required for intranuclear targeting of 53BP1, yet makes no discernable contribution to 53BP1 nuclear import. Of note, depletion of a third nucleoporin (Tpr), which is part of the same structural element of the nuclear pore, did not alter this DNA damage response pathway. To gain insight into the pathway affected by Nup153 and Nup50 and to understand how their role fits into a clinically relevant context, we probed the consequences of reducing Nup153 and Nup50 levels in cells deficient in BRCA1 activity. We found that the requirement for Nup153 and Nup50 in promoting 53BP1 recruitment to damage foci is abrogated under these circumstances. In cells where HR repair is otherwise defective, however, robust recruitment of 53BP1 to DNA damage foci requires Nup153 and Nup50. Together, our results provide novel insight into 53BP1 regulation during the DNA damage response and are consistent with a model where Nup153 and Nup50 normally promote 53BP1 targeting by counteracting BRCA1-dependent events. These findings have important implications with regard to PARP inhibition, a therapeutic strategy thought to capitalize on defective HR repair by forcing inappropriate use of NHEJ, culminating in cellular toxicity. Accordingly, we found that decreased levels of Nup153 and Nup50 do not affect the 53BP1-dependent pathway in BRCA1-deficient cells exposed to PARP inhibitor treatment, but still impair this response in cells deficient for BRCA2 or when the ATM and ATR signaling pathways are inhibited. Our results indicate that Nup153 and Nup50, as factors that modulate 53BP1 activity, belong to a novel category of biomarkers that could be screened to make better predictions of response to PARP inhibitor treatment. Citation Format: Douglas R. Mackay, Katharine S. Ullman. Nup153 and Nup50 promote recruitment of 53BP1 to DNA repair foci by antagonizing BRCA1-dependent events [abstract]. In: Proceedings of the AACR Special Conference on DNA Repair: Tumor Development and Therapeutic Response; 2016 Nov 2-5; Montreal, QC, Canada. Philadelphia (PA): AACR; Mol Cancer Res 2017;15(4_Suppl):Abstract nr A46.

New strategies for the expression of human P‐glycoprotein (MDR1) in the yeast Pichia pastoris

P‐glycoprotein (P‐gp) overexpression is often correlated with multidrug‐resistance (MDR) of cancer. The search for P‐gp inhibitors as co‐therapeutics to combat MDR has had little success after decades of studies. We previously used high‐throughput in silico ligand docking studies and identified drug‐like compounds that reversed MDR in human cancer cells in culture. Effects on P‐gp ATPase hydrolysis activity was used as an initial biochemical screen for inhibitor effectiveness followed by assays of MDR‐reversal of a prostate cancer cell line ((Brewer et al. in Mol Pharmacol. 2014; Follit et al in Pharmacol Res Perspect. 2015). More recently, computationally identified compounds were shown to reverse MDR in cancer cells, but did not appear to inhibit ATP hydrolysis by isolated P‐gp. One reason may be that mouse P‐gp rather than human P‐gp was previously used in our biochemical tests. Expression of human P‐gp in Pichia pastoris in a biologically active form was previously reported, but unfortunately, we experienced very modest expression levels of the human P‐gp expression construct for P. pastoris available in our lab. Codon optimization of mouse P‐gp was previously shown by others to result in increased protein yield, therefore a gene for human P‐gp was designed with optimized codon usage for translation in P. pastoris. A strong Kozak sequence was incorporated at the 5′ end of the coding frame. Unique internal restriction sites were incorporated that allow subcloning of individual sections of the gene, if necessary for troubleshooting and future mutagenesis. The DNA was synthesized using standard Gibson assembly methods and cloned into a pUC57 cloning vector. When transformed into E. coli, DNA rearrangement was observed. To identify whether a specific section of the insert was the cause of counter‐selection in E. coli, the insert DNA was digested into 5 component pieces and each one was subcloned. One of the sections was found to be more susceptible to rearrangements. We report here our attempts at constructing a stable, codon‐optimized, human P‐gp gene for high yield protein production in Pichia pastoris.Support or Funding InformationThis work is supported by NIH NIGMS [R15GM094771‐02] to JGW/PDV, SMU University Research Council, SMU Engaged Learning program, the SMU Center for Drug Discovery, Design and Delivery, the Communities Foundation of Texas, and a private gift from Ms. Suzy Ruff of Dallas, Texas.

Is it time to retire the genus Rymovirus from the family Potyviridae?

In the most recent Report of the International Committee on Taxonomy of Viruses (9th Report, 2011) (King et al., Virus Taxonomy, Elsevier, New York, 2011) the family Potyviridae is described as comprising seven genera - Potyvirus, Ipomovirus, Macluravirus, Rymovirus, Tritimovirus, Brambyvirus and Bymovirus - despite previous suggestions questioning the validity of the taxonomic status of the genus Rymovirus. Since then the ICTV website records that an eighth genus Poacevirus has been approved for the Potyviridae family. The creation of the genus Rymovirus at the 1990 Potyvirus Taxonomy Workshop in Braunschweig, Germany was based on two things: (i) the incorrect assumption that the genomes of all mite-transmitted members of the Potyviridae would have strong sequence similarity to that of wheat streak mosaic virus, the only mite-transmitted member of this genus for which sequence data were available at that time, and (ii) that the genus should be named Rymovirus (based on a virus for which there was no sequence information) rather than a name based on wheat streak mosaic virus (e.g., "Whestremovirus") because ryegrass mosaic virus (RGMV) was the first mite-transmitted virus to be described and thus should take precedence. When sequence data for RGMV became available in 1995, these data showed that RGMV was very different from wheat streak mosaic virus (WSMV) and should not be assigned to the same genus. WSMV was subsequently re-assigned to a new genus, Tritimovirus, while the genus Rymovirus was retained. In this author's opinion, this retention is not justified, and the removal of Rymovirus as a distinct genus in the family Potyviridae is recommended. There may be merit when assigning it to the genus Potyvirus in sequestering these viruses in a rymovirus subgroup, as is done with other potyviruses, to reflect their different mode of transmission.

A putative multi-replicon plasmid co-harboring beta-lactamase genes blaKPC-2, blaCTX-M-14 and blaTEM-1 and trimethoprim resistance gene dfrA25 from a Klebsiella pneumoniae sequence type (ST) 11 strain in China.

The global emergence of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae poses a major public health threat requiring immediate and aggressive action. Some older generation antibiotics, such as trimethoprim, serve as alternatives for treatment of infections. Here, we determined the complete nucleotide sequence of plasmid pHS091147, which co-harbored the carbapenemase (blaKPC-2) and trimethoprim resistance genes (dfrA25) from a Klebsiella pneumoniae sequence type (ST) 11 clone recovered in Shanghai, China. pHS091147 had three replication genes, several plasmid-stability genes and an intact type IV secretion system gene cluster. Besides blaKPC-2 and dfrA25, pHS091147 carried several other resistance genes, including β-lactamase genes blaTEM-1 and blaCTX-M-14, sulphonamide resistance gene sul1, a quinolone resistance gene remnant (ΔqnrB2), and virulence associated gene iroN. Notably, the multidrug-resistance region was a chimeric structure composed of three subregions, which shared strong sequence homology with several plasmids previously assigned in Genbank. To our knowledge, this is the first report of the co-localization of blaKPC-2 and dfrA25 on a novel putative multi-replicon plasmid in a Klebsiella pneumoniae ST11 clone.

Identifying the plant-associated microbiome across aquatic and terrestrial environments: the effects of amplification method on taxa discovery.

Plants in terrestrial and aquatic environments contain a diverse microbiome. Yet, the chloroplast and mitochondria organelles of the plant eukaryotic cell originate from free-living cyanobacteria and Rickettsiales. This represents a challenge for sequencing the plant microbiome with universal primers, as ~99% of 16S rRNA sequences may consist of chloroplast and mitochondrial sequences. Peptide nucleic acid clamps offer a potential solution by blocking amplification of host-associated sequences. We assessed the efficacy of chloroplast and mitochondria-blocking clamps against a range of microbial taxa from soil, freshwater and marine environments. While we found that the mitochondrial blocking clamps appear to be a robust method for assessing animal-associated microbiota, Proteobacterial 16S rRNA binds to the chloroplast-blocking clamp, resulting in a strong sequencing bias against this group. We attribute this bias to a conserved 14-bp sequence in the Proteobacteria that matches the 17-bp chloroplast-blocking clamp sequence. By scanning the Greengenes database, we provide a reference list of nearly 1500 taxa that contain this 14-bp sequence, including 48 families such as the Rhodobacteraceae, Phyllobacteriaceae, Rhizobiaceae, Kiloniellaceae and Caulobacteraceae. To determine where these taxa are found in nature, we mapped this taxa reference list against the Earth Microbiome Project database. These taxa are abundant in a variety of environments, particularly aquatic and semiaquatic freshwater and marine habitats. To facilitate informed decisions on effective use of organelle-blocking clamps, we provide a searchable database of microbial taxa in the Greengenes and Silva databases matching various n-mer oligonucleotides of each PNA sequence.

The effectiveness of DNA‐based delimitation inSynchonnusnet‐winged beetles (Coleoptera: Lycidae) assessed, and description of 11 new species

AbstractSynchonnusWaterhouse, 1879 from Australia is revised.AchrasWaterhouse, 1879 andEnylusWaterhouse, 1879 are found to be junior synonyms ofSynchonnusdue to the absence of diagnostic characters and result in the new combinations ofSynchonnus amplus(Kleine, 1930),S. limbatum(Waterhouse, 1877), andS. segregatus(Waterhouse, 1879).Synchonnusis reported from the Australian mesic and monsoon zones, and 11 new species are described:S. flavonotatumsp. nov.,S. masekisp. nov.,S. ailaketoaesp. nov.,S. dubenovaesp. nov.,S. chilvertonensissp. nov.,S. slipinskiisp. nov.,S. monteithisp. nov.,S. eungellensissp. nov.,S. crypticumsp. nov.,S. variabilissp. nov. andS. campestrissp. nov. The morphology‐based species limits are compared with delimitation inferred from the shape of the phylogenetic tree and genetic distance. DNA‐based species limits agree with morphological delimitation in two clades, but a deep conflict was identified in another clade ofSynchonnusconsisting of three species with allopatric distributions and diversified genitalia, but strong similarities incox1mtDNA sequences. The failure of molecular species delimitation in someSynchonnuspoints to our inability to predict the performance of a barcoding approach even in closely related lineages and calls for an integrative taxonomical approach whenever possible. TheSynchonnusfauna of Australia is presented as highly diverse and fragmentation of habitat in the last ~15 million years is hypothesised as the principal factor leading to the observed alpha‐taxonomic diversity.

Magical Musical Tour: Rock and Pop in Film Soundtracks by K. J. Donnelly

Magical Musical Tour: Rock and Pop in Film Soundtracks. By K. J. Donnelly. New York: Bloomsbury Academic, 2015. [vii, 207 p. ISBN 9781628921281 (hardcover), $100; ISBN 9781628927481 (paperback), $29.95; ISBN 9781628927146 (e-book), $25.99.] Notes, bibliography, index.While recent scholarship has given due individual focus to film soundtracks, rock 'n' roll, and popular music, a volume examining their intersection is certainly a welcome addition to the canon. K. J. (Kevin) Donnelly, a Reader in Film at the University of Southampton, discusses several areas in this vein; overall, the book is a cleanly written survey of topics that (in this reviewer's experience) are all generally familiar, yet Donnelly regularly includes a sub-discussion or reference that entices the reader to pursue further study.The book's introduction provides an overview of some larger concepts within the discourse of film soundtracks, including the basic notion that and film images should not be thought of as two distinct areas, but rather a merged entity. Hollywoodproduced musicals reached an apex in the late 1940s through the early 1950s, accompanied by a decline in the sales of songs, in direct contrast to the emergence of rock 'n' roll records. The success in the 1960s of Cliff Richard's films in England and beach movies in the United States eventually gave way to an art mode of (p. 5) that transitioned rock away from its status as a niche genre toward recognition as an official branch of pop music. By the 1970s, rock was considered different from show business, and the evolving counterculture served as impetus for rock documentaries, with rock musicals and operas solidifying that artistic status. Pairing films with pop songs became de rigueur for marketing in the 1980s, tapping into younger viewers with high school movies and expanding techniques learned from MTV; the 1990s saw more films concerned with rock's (relatively recent) past, and the early twenty-first century enjoyed a renaissance of dance movies.Donnelly spends some time addressing subtler issues (such as the advent of the video) that rejuvenated (and video), while noting how rock and pop are epitomized by the strain between creativity and commerce. While studio technologies behind the production of classical musicals resulted in idealized products grounded in recognizable sequences and using wellknown actors, creators of pop and rock musicals were challenged with negotiating the dual modes of realtime performance (where on-screen images the simultaneous production of sounds being heard) with lip synching. Citing the work of Rick Altman (p. 11), Donnelly discusses the concept of audio dissolve and how the emergence of into the foreground-a development that influenced movements and images, much the opposite of traditional filmmaking-combined with audiovisual aesthetics to create a convergence of pop and rock with traditional film scoring. This development tends toward a lack of subtlety, with emphasis on drumbeats and a domination of sound over image as it becomes more autonomous from pictures on the screen.Donnelly devotes each chapter to an artist or trend within pop film soundtracks, beginning logically with The Beatles. For movies that followed, A Hard Day's Night provided several models, including focus on live performance, a parade of personalities for the artists, use of a strong title sequence, and a climactic conclusion (in this case, the concert broadcast). Help! offered the distinct additions of color film and exotic locations; however, the movie was still constructed around framing the songs. Donnelly touches on two of the film's most memorable sequences: the skiing music video for Ticket to Ride, and the outdoor recording session (with military protection) on Salisbury Plain. Both films share the tension between creation of a dramatic narrative, in which the band members evolve from simple pop stars to songwriters composing for their own art, to simple showcasing of the band and its music. …

Striking Diversity in Holoenzyme Architecture and Extensive Conformational Variability in Biotin-Dependent Carboxylases.

Biotin-dependent carboxylases are widely distributed in nature and have central roles in the metabolism of fatty acids, amino acids, carbohydrates, and other compounds. The last decade has seen the accumulation of structural information on most of these large holoenzymes, including the 500-kDa dimeric yeast acetyl-CoA carboxylase, the 750-kDa α6β6 dodecameric bacterial propionyl-CoA carboxylase, 3-methylcrotonyl-CoA carboxylase, and geranyl-CoA carboxylase, the 720-kDa hexameric bacterial long-chain acyl-CoA carboxylase, the 500-kDa tetrameric bacterial single-chain pyruvate carboxylase, the 370-kDa α2β4 bacterial two-subunit pyruvate carboxylase, and the 130-kDa monomeric eukaryotic urea carboxylase. A common theme that has emerged from these studies is the dramatic structural flexibility of these holoenzymes despite their strong overall sequence conservation, evidenced both by the extensive diversity in the architectures of the holoenzymes and by the extensive conformational variability of their domains and subunits. This structural flexibility is crucial for the function and regulation of these enzymes and identifying compounds that can interfere with it represents an attractive approach for developing novel modulators and drugs. The extensive diversity observed in the structures so far and its biochemical and functional implications will be the focus of this review.

An Update on P2Y13 Receptor Signalling and Function.

The distribution of nucleotide P2Y receptors across different tissues suggests that they fulfil key roles in a number of physiological and pathological conditions. P2Y13 is one of the latest P2Y receptors identified, a novel member of the Gi-coupled P2Y receptor subfamily that responds to ADP, together with P2Y12 and P2Y14. Pharmacological studies drew attention to this new ADP receptor, with a pharmacology that overlaps that of P2Y12 receptors but with unique features and roles. The P2RY12-14 genes all reside on human chromosome 3 at 3q25.1 and their strong sequence homology supports their evolutionary origin through gene duplication. Polymorphisms of P2Y13 receptors have been reported in different human populations, yet their consequences remain unknown. The P2Y13 receptor is versatile in its signalling, extending beyond the canonical signalling of a Gi-coupled receptor. Not only can it couple to different G proteins (Gs/Gq) but the P2Y13 receptor can also trigger several intracellular pathways related to the activation of MAPKs (mitogen-activated protein kinases) and the phosphatidylinositol 3-kinase/Akt/glycogen synthase kinase 3 axis. Moreover, the availability of P2Y13 receptor knockout mice has highlighted the specific functions in which it is involved, mainly in the regulation of cholesterol and glucose metabolism, bone homeostasis and aspects of central nervous system function like pain transmission and neuroprotection. This review summarizes our current understanding of this elusive receptor, not only at the pharmacological and molecular level but also, in terms of its signalling properties and specific functions, helping to clarify the involvement of P2Y13 receptors in pathological situations.