Setting the research agenda in IHRM

Firstly, on behalf of the secretariat that has coordinated these meetings every two years since 1985, our thanks to the organising committee here at the University of Economics in Cracow, Poland, for hosting this conference. I was asked to offer comment on the research agenda. There are many famous names to refer to. Two Australian colleagues here today are Peter Dowling and Helen De Cieri, longtime stalwarts of the field of IHRM. I acknowledge their contributions over many years, along with Randy Schuler and Denise Welch, and Dennis Briscoe. Other names such as Rosalie Tung, Pawan Bhudwhar, Michael Morley, Paul Sparrow and Wayne Cascio are known to us all. Their books have become classics. One example is the 700 page benchmark 2012 work by Chris Brewster and Wolfgang Mayrhofer, Handbook of Research on Comparative Human Resource Management (Brewster & Mayrhofer, 2012). More recently, in a book published by Cambridge University press in 2014, Mustafa Özbilgin, Dimitria Groutsis and William Harvey offer students a very accessible overview of the basics in IHRM (Ozbilgin, Groutsis, & Harvey, 2014). As for a research agenda, there are excellent literature reviews to which I would refer you, such as those by people who over the years have been frequent participants at this conference (Tarique & Schuler, 2010), (Farndale, Scullion, & Sparrow, 2010), and (Scullion & Collings, 2011).

Geminin and Brahma act antagonistically to regulate EGFR–Ras–MAPK signaling in Drosophila

Geminin was identified in Xenopus as a dual function protein involved in the regulation of DNA replication and neural differentiation. In Xenopus, Geminin acts to antagonize the Brahma (Brm) chromatin-remodeling protein, Brg1, during neural differentiation. Here, we investigate the interaction of Geminin with the Brm complex during Drosophila development. We demonstrate that Drosophila Geminin (Gem) interacts antagonistically with the Brm–BAP complex during wing development. Moreover, we show in vivo during wing development and biochemically that Brm acts to promote EGFR–Ras–MAPK signaling, as indicated by its effects on pERK levels, while Gem opposes this. Furthermore, gem and brm alleles modulate the wing phenotype of a Raf gain-of-function mutant and the eye phenotype of a EGFR gain-of-function mutant. Western analysis revealed that Gem over-expression in a background compromised for Brm function reduces Mek (MAPKK/Sor) protein levels, consistent with the decrease in ERK activation observed. Taken together, our results show that Gem and Brm act antagonistically to modulate the EGFR–Ras–MAPK signaling pathway, by affecting Mek levels during Drosophila development.

Molecular basis for lysine specificity in the yeast ubiquitin-conjugating enzyme Cdc34

Ubiquitin (Ub)-conjugating enzymes (E2s) and ubiquitin ligases (E3s) catalyze the attachment of Ub to lysine residues in substrates and Ub during monoubiquitination and polyubiquitination. Lysine selection is important for the generation of diverse substrate-Ub structures, which provides versatility to this pathway in the targeting of proteins to different fates. The mechanisms of lysine selection remain poorly understood, with previous studies suggesting that the ubiquitination site(s) is selected by the E2/E3-mediated positioning of a lysine(s) toward the E2/E3 active site. By studying the polyubiquitination of Sic1 by the E2 protein Cdc34 and the RING E3 Skp1/Cul1/F-box (SCF) protein, we now demonstrate that in addition to E2/E3-mediated positioning, proximal amino acids surrounding the lysine residues in Sic1 and Ub are critical for ubiquitination. This mechanism is linked to key residues composing the catalytic core of Cdc34 and independent of SCF. Changes to these core residues altered the lysine preference of Cdc34 and specified whether this enzyme monoubiquitinated or polyubiquitinated Sic1. These new findings indicate that compatibility between amino acids surrounding acceptor lysine residues and key amino acids in the catalytic core of ubiquitin-conjugating enzymes is an important mechanism for lysine selection during ubiquitination.

Control of cell cycle progression by phosphorylation of cyclin-dependent kinase (CDK) substrates

The eukaryotic cell cycle is a fundamental evolutionarily conserved process that regulates cell division from simple unicellular organisms, such as yeast, through to higher multicellular organisms, such as humans. The cell cycle comprises several phases, including the S-phase (DNA synthesis phase) and M-phase (mitotic phase). During S-phase, the genetic material is replicated, and is then segregated into two identical daughter cells following mitotic M-phase and cytokinesis. The S- and M-phases are separated by two gap phases (G1 and G2) that govern the readiness of cells to enter S- or M-phase. Genetic and biochemical studies demonstrate that cell division in eukaryotes is mediated by CDKs (cyclin-dependent kinases). Active CDKs comprise a protein kinase subunit whose catalytic activity is dependent on association with a regulatory cyclin subunit. Cell-cycle-stage-dependent accumulation and proteolytic degradation of different cyclin subunits regulates their association with CDKs to control different stages of cell division. CDKs promote cell cycle progression by phosphorylating critical downstream substrates to alter their activity. Here, we will review some of the well-characterized CDK substrates to provide mechanistic insights into how these kinases control different stages of cell division.

An inordinate fondness for Fusarium: Phylogenetic diversity of fusaria cultivated by ambrosia beetles in the genus Euwallacea on avocado and other plant hosts

Ambrosia beetle fungiculture represents one of the most ecologically and evolutionarily successful symbioses, as evidenced by the 11 independent origins and 3500 species of ambrosia beetles. Here we document the evolution of a clade within Fusarium associated with ambrosia beetles in the genus Euwallacea (Coleoptera: Scolytinae). Ambrosia Fusarium Clade (AFC) symbionts are unusual in that some are plant pathogens that cause significant damage in naive natural and cultivated ecosystems, and currently threaten avocado production in the United States, Israel and Australia. Most AFC fusaria produce unusual clavate macroconidia that serve as a putative food source for their insect mutualists. AFC symbionts were abundant in the heads of four Euwallacea spp., which suggests that they are transported within and from the natal gallery in mandibular mycangia. In a four-locus phylogenetic analysis, the AFC was resolved in a strongly supported monophyletic group within the previously described Cade 3 of the Fusarium solani species complex (FSSC). Divergence-time estimates place the origin of the AFC in the early Miocene similar to 21.2 Mya, which coincides with the hypothesized adaptive radiation of the Xyleborini. Two strongly supported clades within the AFC (Clades A and B) were identified that include nine species lineages associated with ambrosia beetles, eight with Euwallacea spp. and one reportedly with Xyleborus ferrugineus, and two lineages with no known beetle association. More derived lineages within the AFC showed fixation of the clavate (club-shaped) macroconidial trait, while basal lineages showed a mix of clavate and more typical fusiform macroconidia. AFC lineages consisted mostly of genetically identical individuals associated with specific insect hosts in defined geographic locations, with at least three interspecific hybridization events inferred based on discordant placement in individual gene genealogies and detection of recombinant loci. Overall, these data are consistent with a strong evolutionary trend toward obligate symbiosis coupled with secondary contact and interspecific hybridization. (C) 2013 Elsevier Inc. All rights reserved.

Resent radar observations of asteroid 1566 Icarus

We report Doppler-only radar observations of Icarus at Goldstone at a transmitter frequency of 8510 MHz (3.5 cm wavelength) during 8-10 June 1996, the first radar detection of the object since 1968. Optimally filtered and folded spectra achieve a maximum opposite-circular (OC) polarization signal-to-noise ratio of about 10 and help to constrain Icarus' physical properties. We obtain an OC radar cross section of 0.05 km(2) (with a 35% uncertainty), which is less than values estimated by Goldstein (1969) and by Pettengill et al. (1969), and a circular polarization (SC/OC) ratio of 0.5+/-0.2. We analyze the echo power spectrum with a model incorporating the echo bandwidth B and a spectral shape parameter it, yielding a coupled constraint between B and n. We adopt 25 Hz as the lower bound on B, which gives a lower bound on the maximum pole-on breadth of about 0.6 km and upper bounds on the radar and optical albedos that are consistent with Icarus' tentative QS classification. The observed circular polarization ratio indicates a very rough near-surface at spatial scales of the order of the radar wavelength. (C) 1999 Elsevier Science Ltd. All rights reserved.

Novel Pentameric Structure of the Diarrhea-Inducing Region of the Rotavirus Enterotoxigenic Protein NSP4

A novel pentameric structure which differs from the previously reported tetrameric form of the diarrhea-inducing region of the rotavirus enterotoxin NSP4 is reported here. A significant feature of this pentameric form is the absence of the calcium ion located in the core region of the tetrameric structures. The lysis of cells, the crystallization of the region spanning residues 95 to 146 of NSP4 (NSP4(95-146)) of strain ST3 (ST3: NSP4(95-146)) at acidic pH, and comparative studies of the recombinant purified peptide under different conditions by size-exclusion chromatography (SEC) and of the crystal structures suggested pH-, Ca(2+)-, and protein concentration-dependent oligomeric transitions in the peptide. Since the NSP4(95-146) mutant lacks the N-terminal amphipathic domain (AD) and most of the C-terminal flexible region (FR), to demonstrate that the pentameric transition is not a consequence of the lack of the N- and C-terminal regions, glutaraldehyde cross-linking of the Delta N72 and Delta N94 mutant proteins, which contain or lack the AD, respectively, but possess the complete C-terminal FR, was carried out. The results indicate the presence of pentamers in preparations of these longer mutants. Detailed SEC analyses of Delta N94 prepared under different conditions, however, revealed protein concentration-dependent but metal ion-and pH-independent pentamer accumulation at high concentrations which dissociated into tetramers and lower oligomers at low protein concentrations. While calcium appeared to stabilize the tetramer, magnesium in particular stabilized the dimer. Delta N72 existed primarily in the multimeric form under all conditions. These findings of a calcium-free NSP4 pentamer and its concentration-dependent and largely calcium-independent oligomeric transitions open up a new dimension in an understanding of the structural basis of its multitude of functions.

A new pentameric structure of rotavirus NSP4 revealed by molecular replacement

The region spanning residues 95-146 of the rotavirus nonstructural protein NSP4 from the asymptomatic human strain ST3 has been purified and crystallized and diffraction data have been collected to a resolution of 2.6 angstrom. Several attempts to solve the structure by the molecular-replacement method using the available tetrameric structures of this domain were unsuccessful despite a sequence identity of 73% to the already known structures. A more systematic approach with a dimer as the search model led to an unexpected pentameric structure using the program Phaser. The various steps involved in arriving at this molecular-replacement solution, which unravelled a case of subtle variation between different oligomeric states unknown at the time of solving the structure, are presented in this paper.

Severe diffraction anisotropy, rotational pseudosymmetry and twinning complicate the refinement of a pentameric coiled-coil structure of NSP4 of rotavirus

The crystal structure of the region spanning residues 95-146 of the rotavirus nonstructural protein NSP4 from the asymptomatic human strain ST3 was determined at a resolution of 2.5 angstrom. Severe diffraction anisotropy, rotational pseudo-symmetry and twinning complicated the refinement of this structure. A systematic explanation confirming the crystal pathologies and describing how the structure was successfully refined is given in this report.

Engineering enzyme systems by recombination

Homologous recombination is a source of diversity in both natural and directed evolution. Standing genetic variation that has passed the test of natural selection is combined in new ways, generating functional and sometimes unexpected changes. In this work we evaluate the utility of homologous recombination as a protein engineering tool, both in comparison with and combined with other protein engineering techniques, and apply it to an industrially important enzyme: Hypocrea jecorina Cel5a.

Chapter 1 reviews work over the last five years on protein engineering by recombination. Chapter 2 describes the recombination of Hypocrea jecorina Cel5a endoglucanase with homologous enzymes in order to improve its activity at high temperatures. A chimeric Cel5a that is 10.1 °C more stable than wild-type and hydrolyzes 25% more cellulose at elevated temperatures is reported. Chapter 3 describes an investigation into the synergy of thermostable cellulases that have been engineered by recombination and other methods. An engineered endoglucanase and two engineered cellobiohydrolases synergistically hydrolyzed cellulose at high temperatures, releasing over 200% more reducing sugars over 60 h at their optimal mixture relative to the best mixture of wild-type enzymes. These results provide a framework for engineering cellulolytic enzyme mixtures for the industrial conditions of high temperatures and long incubation times.

In addition to this work on recombination, we explored three other problems in protein engineering. Chapter 4 describes an investigation into replacing enzymes with complex cofactors with simple cofactors, using an E. coli enolase as a model system. Chapter 5 describes engineering broad-spectrum aldehyde resistance in Saccharomyces cerevisiae by evolving an alcohol dehydrogenase simultaneously for activity and promiscuity. Chapter 6 describes an attempt to engineer gene-targeted hypermutagenesis into E. coli to facilitate continuous in vivo selection systems.

A comparison between circle hook and J hook performance in the dolphinfish, yellowfish tuna, and wahoo troll fishery off the coast of North Carolina

We compared numbers of strikes, proportions of fish that hooked up after strikes, proportions of fish that stayed on hook (retained) after hook up, and numbers of fish caught between circle and J hooks rigged with dead natural fish bait (ballyhoo)and trolled for three oceanic predator species: dolphinfish (Coryphaena hippurus), yellowfin tuna (Thunnus albacares), and wahoo (Acanthocybium solandri). Interactions were compared between circle and J hooks fished on 75 trips by two user groups (charter and recreational fishermen). Hooks were affixed to three species-specific leader types most commonly fished in this region: monofilament (dolphinfish), fluorocarbon (tuna), and wire (wahoo). Numbers of fish caught per trip and three potential mechanisms that might inf luence numbers caught (i.e., number of strikes, proportion of fish hooked, and proportion retained) were modeled with generalized linear models that considered hook type, leader type, species, user (fishing) group, and wave height as main effects. Hook type was a main effect at the catch level; generally, more fish were caught on J hooks than on circle hooks. The effect of hook type on strike rates was equivocal. However, J hooks had a greater proportion of hook-ups than did circle hooks. Finally, the proportion of fish retained once hooked was generally equal between hook types. We found similar results when data from additional species were pooled as a “tuna” group and a “mackerel” group. We conclude that J hooks are more effective than circle hooks at the hook-up level and result in greater numbers of troll-caught dolphinfish, tunas

An ecological characterization of the Stellwagen Bank National Marine Sanctuary Region: oceanographic, biogeographic, and contaminants assessment

The mission of NOAA’s National Marine Sanctuary Program (NMSP) is to serve as the trustee for a system of marine protected areas, to conserve, protect, and enhance their biodiversity, ecological integrity, and cultural legacy while facilitating compatible uses. Since 1972, thirteen National Marine Sanctuaries, representing a wide variety of ocean environments, have been established, each with management goals tuned to their unique diversity. Extending from Cape Ann to Cape Cod across the mouth of Massachusetts Bay, Stellwagen Bank National Marine Sanctuary (NMS) encompasses 2,181 square kilometers of highly productive, diverse, and culturally unique Federal waters. As a result of its varied seafloor topography, oceanographic conditions, and high primary productivity, Stellwagen Bank NMS is utilized by diverse assemblages of seabirds, marine mammals, invertebrates, and fish species, as well as containing a number of maritime heritage resources. Furthermore, it is a region of cultural significance, highlighted by the recent discovery of several historic shipwrecks. Officially designated in 1992, Stellwagen Bank became the Nation’s twelfth National Marine Sanctuary in order to protect these and other unique biological, geological, oceanographic, and cultural features of the region. The Stellwagen Bank NMS is in the midst of its first management plan review since designation. The management plan review process, required by law, is designed to evaluate, enhance, and guide the development of future research efforts, education and outreach, and the management approaches used by Sanctuaries. Given the ecological and physical complexity of Stellwagen Bank NMS, burgeoning anthropogenic impacts to the region, and competing human and biological uses, the review process was challenged to assimilate and analyze the wealth of existing scientific knowledge in a framework which could enhance management decision-making. Unquestionably, the Gulf of Maine, Massachusetts Bay, and Stellwagen Bank-proper are extremely well studied systems, and in many regards, the scientific information available greatly exceeds that which is available for other Sanctuaries. However, the propensity of scientific information reinforces the need to utilize a comprehensive analytical approach to synthesize and explore linkages between disparate information on physical, biological, and chemical processes, while identifying topics needing further study. Given this requirement, a partnership was established between NOAA’s National Marine Sanctuary Program (NMSP) and the National Centers for Coastal Ocean Science (NCCOS) so as to leverage existing NOAA technical expertise to assist the Sanctuary in developing additional ecological assessment products which would benefit the management plan review process.

A biogeographic assessment of the Channel Islands National Marine Sanctuary: a review of boundary expansion concepts for NOAA’s National Marine Sanctuary Program

The priority management goal of the National Marine Sanctuaries Program (NMSP) is to protect marine ecosystems and biodiversity. This goal requires an understanding of broad-scale ecological relationships and linkages between marine resources and physical oceanography to support an ecosystem management approach. The Channel Islands National Marine Sanctuary (CINMS) is currently reviewing its management plan and investigating boundary expansion. A management plan study area (henceforth, Study Area) was described that extends from the current boundary north to the mainland, and extends north to Point Sal and south to Point Dume. Six additional boundary concepts were developed that vary in area and include the majority of the Study Area. The NMSP and CINMS partnered with NOAA’s National Centers for Coastal Ocean Science Biogeography Team to conduct a biogeographic assessment to characterize marine resources and oceanographic patterns within and adjacent to the sanctuary. This assessment includes a suite of quantitative spatial and statistical analyses that characterize biological and oceanographic patterns in the marine region from Point Sal to the U.S.-Mexico border. These data were analyzed using an index which evaluates an ecological “cost-benefit” within the proposed boundary concepts and the Study Area. The sanctuary resides in a dynamic setting where two oceanographic regimes meet. Cold northern waters mix with warm southern waters around the Channel Islands creating an area of transition that strongly influences the regions oceanography. In turn, these processes drive the biological distributions within the region. This assessment analyzes bathymetry, benthic substrate, bathymetric life-zones, sea surface temperature, primary production, currents, submerged aquatic vegetation, and kelp in the context of broad-scale patterns and relative to the proposed boundary concepts and the Study Area. Boundary cost-benefit results for these parameters were variable due to their dynamic nature; however, when analyzed in composite the Study Area and Boundary Concept 2 were considered the most favorable. Biological data were collected from numerous resource agencies and university scientists for this assessment. Fish and invertebrate trawl data were used to characterize community structure. Habitat suitability models were developed for 15 species of macroinvertebrates and 11 species of fish that have significant ecological, commercial, or recreational importance in the region and general patterns of ichthyoplankton distribution are described. Six surveys of ship and plane at-sea surveys were used to model marine bird diversity from Point Arena to the U.S.-Mexico border. Additional surveys were utilized to estimate density and colony counts for nine bird species. Critical habitat for western snowy plover and the location of California least tern breeding pairs were also analyzed. At-sea surveys were also used to describe the distribution of 14 species of cetaceans and five species of pinnipeds. Boundary concept cost-benefit indices revealed that Boundary Concept 2 and the Study Area were most favorable for the majority of the species-specific analyses. Boundary Concept 3 was most favorable for bird diversity across the region. Inadequate spatial resolution for fish and invertebrate community data and incompatible sampling effort information for bird and mammal data precluded boundary cost-benefit analysis.

Survey and impact assessment of derelict fish traps in St. Thomas and St. John, U.S. Virgin Islands

Since 2001, NOAA National Centers for Coastal Ocean Science (NCCOS), Center for Coastal Monitoring and Assessment’s (CCMA) Biogeography Branch (BB) has been working with federal and territorial partners to characterize, monitor, and assess the status of the marine environment across the U.S. Virgin Islands (USVI). At the request of the St. Thomas Fisherman’s Association (STFA) and NOAA Marine Debris Program, CCMA BB developed new partnerships and novel technologies to scientifically assess the threat from derelict fish traps (DFTs). Traps are the predominant gear used for finfish and lobster harvesting in St. Thomas and St. John. Natural phenomena (ground swells, hurricanes) and increasing competition for space by numerous user groups have generated concern about increasing trap loss and the possible ecological, as well as economic, ramifications. Prior to this study, there was a general lack of knowledge regarding derelict fish traps in the Caribbean. No spatially explicit information existed regarding fishing effort, abundance and distribution of derelict traps, the rate at which active traps become derelict, or areas that are prone to dereliction. Furthermore, there was only limited information regarding the impacts of derelict traps on natural resources including ghost fishing. This research identified two groups of fishing communities in the region: commercial fishing that is most active in deeper waters (30 m and greater) and an unknown number of unlicensed subsistence and or commercial fishers that fish closer to shore in shallower waters (30 m and less). In the commercial fishery there are an estimated 6,500 active traps (fish and lobster combined). Of those traps, nearly 8% (514) were reported lost during the 2008-2010 period. Causes of loss/dereliction include: movement of the traps or loss of trap markers due to entanglement of lines by passing vessels; theft; severe weather events (storms, large ground swells); intentional disposal by fishermen; traps becoming caught on various bottom structures (natural substrates, wrecks, etc.); and human error.

Fish movement patterns in Virgin Islands National Park, Virgin Islands Coral Reef National Monument and adjacent waters

NOAA’s National Centers for Coastal Ocean Science (NCCOS)-Center for Coastal Monitoring and Assessment’s (CCMA) Biogeography Branch, National Park Service (NPS), US Geological Survey, and the University of Hawaii used acoustic telemetry to quantify spatial patterns and habitat affinities of reef fishes around the island of St. John, US Virgin Islands. The objective of the study was to define the movements of reef fishes among habitats within and between the Virgin Islands Coral Reef National Monument (VICRNM), the Virgin Islands National Park (VIIS), and Territorial waters surrounding St. John. In order to better understand species’ habitat utilization patterns among management regimes, we deployed an array of hydroacoustic receivers and acoustically tagged reef fishes. Thirty six receivers were deployed in shallow near-shore bays and across the shelf to depths of approximately 30 m. One hundred eighty four individual fishes were tagged representing 19 species from 10 different families with VEMCO V9-2L-R64K transmitters. The array provides fish movement information at fine (e.g., day-night and 100s meters within a bay) to broad spatial and temporal scales (multiple years and 1000s meters across the shelf). The long term multi-year tracking project provides direct evidence of connectivity across habitat types in the seascape and among management units. An important finding for management was that a number of individuals moved among management units (VICRNM, VINP, Territorial waters) and several snapper moved from near-shore protected areas to offshore shelf-edge spawning aggregations. However, most individuals spent the majority of their time with VIIS and VICRNM, with only a few wide-ranging species moving outside the management units. Five species of snappers (Lutjanidae) accounted for 31% of all individuals tagged, followed by three species of grunts (Haemulidae) accounting for an additional 23% of the total. No other family had more than a single species represented in the study. Bluestripe grunt (Haemulon sciurus) comprised 22% of all individuals tagged, followed by lane snappers (Lutjanus synagris) at 21%, bar jack (Carangoides ruber) at 11%, and saucereye porgy (Calamus calamus) at 10%. The largest individual tagged was a 70 cm TL nurse shark (Ginglymostoma cirratum), followed by a 65 cm mutton snapper (Lutjanus analis), a 47 cm bar jack, and a 41 cm dog snapper (Lutjanus jocu). The smallest individuals tagged were a 19 cm blue tang (Acanthurus coeruleus) and a 19.2 cm doctorfish (Acanthurus chirurgus). Of the 40 bluestriped grunt acoustically tagged, 73% were detected on the receiver array. The average days at large (DAL) was 249 (just over 8 months), with one individual detected for 930 days (over two and a half years). Lane snapper were the next most abundant species tagged (N = 38) with 89% detected on the array. The average days at large (DAL) was 221 with one individual detected for 351 days. Seventy-one percent of the bar jacks (N = 21) were detected on the array with the average DALs at 47 days. All of the mutton snapper (N = 12) were detected on the array with an average DAL of 273 and the longest at 784. The average maximum distance travelled (MDT) was ca. 2 km with large variations among species. Grunts, snappers, jacks, and porgies showed the greatest movements. Among all individuals across species, there was a positive and significant correlation between size of individuals and MDT and between DAL and MDT.