Re-examining the role of Drosophila Sas-4 in centrosome assembly using two-colour-3D-SIM FRAP.

Centrosomes have many important functions and comprise a 'mother' and 'daughter' centriole surrounded by pericentriolar material (PCM). The mother centriole recruits and organises the PCM and templates the formation of the daughter centriole. It has been reported that several important Drosophila PCM-organising proteins are recruited to centrioles from the cytosol as part of large cytoplasmic 'S-CAP' complexes that contain the centriole protein Sas-4. In a previous paper (Conduit et al., 2014b) we showed that one of these proteins, Cnn, and another key PCM-organising protein, Spd-2, are recruited around the mother centriole before spreading outwards to form a scaffold that supports mitotic PCM assembly; the recruitment of Cnn and Spd-2 is dependent on another S-CAP protein, Asl. We show here, however, that Cnn, Spd-2 and Asl are not recruited to the mother centriole as part of a complex with Sas-4. Thus, PCM recruitment in fly embryos does not appear to require cytosolic S-CAP complexes.PTC was supported by a Sir Henry Dale Fellowship jointly funded by the Wellcome Trust and the Royal Society (105653/Z/14/Z) and by an Issac Newton Trust Research Grant from the University of Cambridge awarded to TTW (RG78799). AW, ZN and JWR were supported by a Senior Investigator Award awarded to JWR and funded by the Wellcome Trust (104575/Z/14/Z). The OMX microscope used in this study is part of the Oxford Micron Advanced Bioimaging Unit supported by a Wellcome Trust Strategic Award (091911).This is the final version of the article. It first appeared from eLife via http://dx.doi.org/10.7554/eLife.0848

Impacts of Venturi Turbulent Mixing on the Size Distribution of Sodium Chloride and Dioctyl-Phthalate Aerosols

Internal combustion engines are a major source of airborne particulate matter (PM). The size of the engine PM is in the sub-micrometer range. The number of engine particles per unit volume is high, normally in the range of 10{sup 12} to 10{sup 14}. To measure the size distribution of the engine particles dilution of an aerosol sample is required. A diluter utilizing a venturi ejector mixing technique is commercially available and tested. The purpose of this investigation was to determine if turbulence created by the ejector in the mini-dilutor changes the size of particles passing through it

Evolution of the Southwest Australian Rifted Continental Margin During Breakup of East Gondwana: Results from IODP Expedition 369

International Ocean Discovery Program Expedition 369 drilled four sites on the southwestern Australian continental margin, in the deep water Mentelle Basin (MB) and on the neighboring Naturaliste Plateau (NP). The drillsites are located on continental crust that continued rifting after seafloor spreading began further north on the Perth Abyssal Plain (PAP) between magnetochrons M11r and M11n (133‐132 Ma), ending when spreading began west of the NP between chrons M5n and M3n (126‐124 Ma). Drilling recovered the first in‐situ samples of basalt flows overlying the breakup unconformity on the NP, establishing a magnetostratigraphically constrained eruption age of >131‐133 Ma and confirming a minimal late Valanginian age for the breakup unconformity (coeval with the onset of PAP seafloor spreading). Petrogenetic modeling indicates the basalts were generated by 25% melting at 1.5 GPa and a potential temperature of 1380‐1410 °C, consistent with proximity of the Kerguelen plume during breakup. Benthic foraminiferal fossils indicate that the NP remained at upper bathyal or shallower depths during the last 6 Myr of rifting and for 3‐5 Myr after breakup between India and Australia. The limited subsidence is attributed to heat from the nearby Kerguelen plume and PAP spreading ridge. The margin subsided to middle bathyal depths by Albian time and to lower bathyal (NP) or greater (MB) depths by late Paleogene time. Periods of rapid sedimentation accompanied a westward jump of the PAP spreading ridge (108 Ma), rifting on the southern margin (100‐84 Ma), and opening of the southern seaway between Australia and Antarctica (60‐47 Ma)

Structural Basis for Mitotic Centrosome Assembly in Flies.

In flies, Centrosomin (Cnn) forms a phosphorylation-dependent scaffold that recruits proteins to the mitotic centrosome, but how Cnn assembles into a scaffold is unclear. We show that scaffold assembly requires conserved leucine zipper (LZ) and Cnn-motif 2 (CM2) domains that co-assemble into a 2:2 complex in vitro. We solve the crystal structure of the LZ:CM2 complex, revealing that both proteins form helical dimers that assemble into an unusual tetramer. A slightly longer version of the LZ can form micron-scale structures with CM2, whose assembly is stimulated by Plk1 phosphorylation in vitro. Mutating individual residues that perturb LZ:CM2 tetramer assembly perturbs the formation of these micron-scale assemblies in vitro and Cnn-scaffold assembly in vivo. Thus, Cnn molecules have an intrinsic ability to form large, LZ:CM2-interaction-dependent assemblies that are critical for mitotic centrosome assembly. These studies provide the first atomic insight into a molecular interaction required for mitotic centrosome assembly.Z.F. and A.F.M.H. were supported by Sir William Dunn School EPA PhD studentships and also a Clarendon Scholarship and a Santander Graduate Award to A.F.M.H; A.C., A.W., M.A.C., P.T.C., and J.W.R. were supported by a Wellcome Trust Senior Investigator Award (104575); S.J. and S.M.L. were supported by a Wellcome Trust Senior Investigator Award (100298); A.W. was also partially supported by a Wellcome Trust Strategic Award to the Micron Oxford Advanced Bioimaging Unit (107457)

Evidence that a positive feedback loop drives centrosome maturation in fly embryos.

Centrosomes are formed when mother centrioles recruit pericentriolar material (PCM) around themselves. The PCM expands dramatically as cells prepare to enter mitosis (a process termed centrosome maturation), but it is unclear how this expansion is achieved. In flies, Spd-2 and Cnn are thought to form a scaffold around the mother centriole that recruits other components of the mitotic PCM, and the Polo-dependent phosphorylation of Cnn at the centrosome is crucial for scaffold assembly. Here, we show that, like Cnn, Spd-2 is specifically phosphorylated at centrosomes. This phosphorylation appears to create multiple phosphorylated S-S/T(p) motifs that allow Spd-2 to recruit Polo to the expanding scaffold. If the ability of Spd-2 to recruit Polo is impaired, the scaffold is initially assembled around the mother centriole, but it cannot expand outwards, and centrosome maturation fails. Our findings suggest that interactions between Spd-2, Polo and Cnn form a positive feedback loop that drives the dramatic expansion of the mitotic PCM in fly embryos

Relations between assemblages of carpological remains and modern vegetation in a shallow reservoir in southern Poland

This paper explores relations between assemblages of carpological remains and vegetation in and around a small, shallow reservoir in southern Poland. The study was conducted from 2006 to 2008. Quantity and distribution of species in the reservoir were recorded annually during the growing season. In October 2008, 40 samples of surface sediment (top 2 cm) were collected along transects at 10 m intervals. Samples of 100 cm3 were prepared for analysis of plant macroremains. Assemblages of carpological remains generally reflect local vegetation well. In some cases, however, even analysis of numerous samples failed to fully capture the species composition or reflect plant ratios in the parent phytocenosis. Reasons for this include factors that affect seed production, transport and fossilization, which differ among species. Among the best-represented macroremains were plants of the rush phytocenosis. In analysed samples, macroremains of 68.8 % of extant rushes were identified. Sixty percent of submerged and floating-leaf taxa were found in carpological samples, whereas 26.7 % of the trees and bushes were represented in sediment deposits. Species composition of phytocenoses in the reservoir and in surrounding areas was best reflected by macroremains from the nearby reed bed. Numbers of diaspores of Mentha aquatica, Hippuris vulgaris and Carex reflected well their relative abundance in phytocenoses. Chara sp., Juncus inflexus and Eupatorium cannabinum were overrepresented, whereas Typha latifolia and Sparganium minimum were poorly represented in relation to contemporary plant cover. There were no diaspores of Phragmites australis, which dominates the contemporary reed bed. Besides the shape of a reservoir, the key factor influencing diaspore numbers is distribution of plant cover. In many cases, single diaspores (Potentilla erecta, Myosotis scorpioides, Lythrum salicaria, Scutellaria galericulata), or higher concentrations (Hippuris vulgaris, Mentha aquatica, Eleocharis palustris, Schoenoplectus tabernaemontani, Chara sp.) reflected well the location of parent vegetation. The findings indicate that carpological remains in sediments can be an important source of information about plants in and around lakes. They generally reflect well local vegetation and in some cases may be used to identify taxa that dominated in the past