TBCD Links Centriologenesis,Spindle Microtubule Dynamics,and Midbody Abscission in Human Cells

Microtubule-organizing centers recruit α- and β-tubulin polypeptides for microtubule nucleation. Tubulin synthesis is complex, requiring five specific cofactors, designated tubulin cofactors (TBCs) A–E, which contribute to various aspects of microtubule dynamics in vivo. Here, we show that tubulin cofactor D (TBCD) is concentrated at the centrosome and midbody, where it participates in centriologenesis, spindle organization, and cell abscission. TBCD exhibits a cell-cycle-specific pattern, localizing on the daughter centriole at G1 and on procentrioles by S, and disappearing from older centrioles at telophase as the protein is recruited to the midbody. Our data show that TBCD overexpression results in microtubule release from the centrosome and G1 arrest, whereas its depletion produces mitotic aberrations and incomplete microtubule retraction at the midbody during cytokinesis. TBCD is recruited to the centriole replication site at the onset of the centrosome duplication cycle. A role in centriologenesis is further supported in differentiating ciliated cells, where TBCD is organized into “centriolar rosettes”. These data suggest that TBCD participates in both canonical and de novo centriolar assembly pathways.     

Interaction between cytochrome c and cytochrome c peroxidase: excited-state reactions of zinc- and tin-substituted derivatives

The effect of cytochrome c peroxidase (CCP) and apoCCP on the fluorescence and phosphorescence of Zn and Sn cytochrome c (cyt c) and the effect of cyt c on the fluorescence and phosphorescence of Zn CCP were examined. We found the following: The fluorescence yields of Zn and Sn cyt c were quenched by about 20% by CCP, consistent with energy transfer between the two chromophores with a separation of about 1.8 nm. The phosphorescence spectrum of Zn cyt c (but not Sn cyt c) shifts by 20 nm to the blue upon complexation with either CCP or apoCCP; at the same time the phosphorescence lifetime of Zn cyt c decreases from 12 +/- 2 to 6 ms with apoCCP addition. Zn CCP phosphorescence decay increases from 8.3 to 9.1 ms upon addition of poly(L-lysine) used to mimic cyt c. It is concluded from these results that binding of the redox partner or an analogue to Zn CCP and Zn cyt c results in a conformational change. The respective phosphorescence lifetimes of Zn and Sn cyt c were 13 and 3 ms in the absence of CCP and 1.6 and 1.1 ms in the presence of CCP; this corresponds to a quenching rate due to CCP of 519 and 570 s-1, for Zn and Sn cyt c, respectively. The phosphorescence of Zn CCP is also affected by native cyt c but is dramatically less than the complementary pair; the quenching rate constant is 17 s-1.(ABSTRACT TRUNCATED AT 250 WORDS)     

Frequencies of simultaneous transformation with different T-DNAs and their relevance to the Agrobacterium/plant cell interaction

Summary We investigated whether the efficiency of transformation of plant cells by Agrobacterium tumefaciens during cocultivation is limited by the properties of the plant cells or by the infecting bacteria.Therefore, tobacco protoplasts were infected by cocultivation with two different agrobacteria strains carrying Ti plasmids with distinguishable T-DNAs. These T-DNAs cotransform plant cells at a frequency equal to the product of their independent transformation frequencies, which indicates that all plant cells are equally competent. On the other hand, when these T-DNAs are located on the same Ti plasmid vector within one bacterial strain, the cotransformation frequency is significantly higher than the product of the single transformation frequencies. We interpret these results to indicate that transformation is limited more by the establishment of effective bacteria/plant cell interaction than by (i) the process of DNA integration and (ii) by the number of plant cells capable of being transformed by Agrobacterium. We found that most plant cells are transformed by only one or a few agrobacteria. Analysis of the number of T-DNA copies in these clonally transformed lines indicates amplification of the original, infecting T-region copy.     

Abundance and biomass estimates of the benthic fauna in Admiralty Bay,King George Island,South Shetland Islands

Summary Quantitative benthic samples were collected along three transects in Admiralty Bay, King George Island, South Shetlands. At each of a total of 18 stations, between 15 and 250 m depth, we took 3 replicate samples with a van Veen grab. Animals collected were classed into major groups. Abundance and biomass per m² was calculated for each sampling site. Considerable population densities and high biomass values were found. Most abundant groups were Bivalvia, Polychaeta and Amphipoda, whereas the largest part of the biomass was due to Ascidiacea, Ophiuroidea, Echinoidea, Polychaeta and Bivalvia. The maximum abundance recorded was 36,000 ind m^-2 while the average was approximately 6500 ind m^-2. Maximum biomass was over 2400 g m^-2 with an average of ca. 700 g m^-2. The contribution to the total biomass by groups such as the Oligochaeta, Cumacea and Tanaidacea was higher in the inner shallow part of Admiralty Bay (Ezcurra Inlet) than in the deeper areas of the bay. Our results confirm the reports on an unusually high density and biomass of the Antarctic sublittoral benthic fauna. Sessile suspension feeders belonging to the Bivalvia, Ascidiacea, sedentary Polychaeta, and vagile scavengers of the Ophiuroidea, Amphipoda and errant Polychaeta are the most significant groups in the Antarctic Ecosystem. The total benthic biomass in Admiralty Bay, based on the present preliminary quantitative data, was estimated to be over 600,000 t. This value is probably still an underestimate.     

Structural characterization of cytochrome c peroxidase by resonance Raman scattering

Resonance Raman scattering studies are reported on freshly prepared and aged ferric, ligand-free ferrous, and CO-bound ferrous cytochrome c peroxidase. The ferric form of the fresh enzyme has a heme which is penta-coordinate high spin, independent of buffer over the pH range 4.3-7, as determined by well established Raman marker lines. The aged enzyme displays a mixture of spin and coordination states, but it can be stabilized in the penta-coordinate high spin form in the presence of phosphate. These results can be accounted for by considering the size of the channel (6 A wide, 11 A long) between the distal side of the heme and the outer surface of the protein. A phosphate ion may be accommodated in this channel resulting in the stabilization of the distal heme pocket. The ferrous cytochrome c peroxidase in both the ligand-free and CO-bound states has an acidic and an alkaline form. The acidic form has the characteristic spectral features of peroxidases: a high frequency iron-histidine stretching mode (248 cm-1), a high frequency Fe-CO stretching mode (537 cm-1), and a low frequency C-O stretching mode (1922 cm-1). At alkaline pH these frequencies become similar to those of hemoglobin and myoglobin, with the corresponding modes located at 227, 510, and 1948 cm-1, respectively. We attribute the acid/alkaline transition in the ferrous forms of cytochrome c peroxidase to a rearrangement mainly of the proximal side of the heme, culminating in a change of steric interactions between the proximal histidine and the heme or of the hydrogen bonding network involving the proximal histidine. The new data presented here reconcile many inconsistencies reported in the past.     

Determination of trace elements in standard reference materials by the ko-standardization method

Biological Trace Element Research - The ko-standardization method is suitable for routine multielement determinations by reactor neutron activation analysis (NAA). Investigation of NIST standard...     

New Zealand machair vegetation

Abstract. Machair vegetation is reported for the first time from New Zealand. The habitat is similar to that of British machairs in climate, topography and generally in soil. pH and CaCO₃ content are much lower through most of the sequence, though this difference may partly reflect the greater disturbance of British machair. Sea machair is present, predominantly comprising native species. This grades into machair proper, which contains many species found also in British machair. The machair includes Ammophila-occupied hillocks, a feature typical of British machair. Machair marsh is also present.     

Lawns have vertical stratification

Abstract. Vertical stratification was examined in a closely-mown lawn, at both the beginning and end of the mowing/regrowth cycle. Two treatments were examined: Undisturbed, which had been maintained as lawn for 30 yr, and Mechanically Perturbed, on which succession was occurring, after complete removal of vegetation 9.5 months previously. There was significant vertical stratification at both stages of the mowing/regrowth cycle, i.e. even immediately after mowing to a height of 3 cm, and in both treatments. There was general consistency of the vertical role of species during the cycle and between treatments. Relative vertical positions were not closely correlated with absolute leaf heights. However, in no case was the vertical position of a pair of species significantly reversed during the cycle or between treatments. No mutually cyclic vertical relations between species were seen. It is concluded that vertical stratification occurs in even the shortest communities, it starts to develop quite early in succession, and it is dependent largely on the intrinsic properties of the species, their height and (at least in lawns) their recovery from defoliation.