Phosphorylation and ubiquitination of dynamin-related proteins (AtDRP3A/3B) synergically regulate mitochondrial proliferation during mitosis

The balance between mitochondrial fission and fusion is disrupted during mitosis, but the mechanism governing this phenomenon in plant cells remains enigmatic. Here, we used mitochondrial matrix‐localized Kaede protein (mt‐Kaede) to analyze the dynamics of mitochondrial fission in BY‐2 suspension cells. Analysis of the photoactivatable fluorescence of mt‐Kaede suggested that the fission process is dominant during mitosis. This finding was confirmed by an electron microscopic analysis of the size distribution of mitochondria in BY‐2 suspension cells at various stages. Cellular proteins interacting with Myc‐tagged dynamin‐related protein 3A/3B (AtDRP3A and AtDRP3B) were immunoprecipitated with anti‐Myc antibody‐conjugated beads and subsequently identified by microcapillary liquid chromatography–quadrupole time‐of‐flight mass spectrometry (CapLC Q‐TOF) MS/MS. The identified proteins were broadly associated with cytoskeletal (microtubular), phosphorylation, or ubiquitination functions. Mitotic phosphorylation of AtDRP3A/AtDRP3B and mitochondrial fission at metaphase were inhibited by treatment of the cells with a CdkB/cyclin B inhibitor or a serine/threonine protein kinase inhibitor. The fate of AtDRP3A/3B during the cell cycle was followed by time‐lapse imaging of the fluorescence of Dendra2‐tagged AtDRP3A/3B after green‐to‐red photoconversion; this experiment showed that AtDRP3A/3B is partially degraded during interphase. Additionally, we found that microtubules are involved in mitochondrial fission during mitosis, and that mitochondria movement to daughter cell was limited as early as metaphase. Taken together, these findings suggest that mitotic phosphorylation of AtDRP3A/3B promotes mitochondrial fission during plant cell mitosis, and that AtDRP3A/3B is partially degraded at interphase, providing mechanistic insight into the mitochondrial morphological changes associated with cell‐cycle transitions in BY‐2 suspension cells.     

Ruptured fission yeast walls

Distributions of rupture sites of fission yeast cells ruptured by glass beads have been related to a new morphometric analysis. As shown previously (Johnson et al.,Cell Biophysics, 1995), ruptures were not randomly distributed nor was their distribution dictated by geometry, rather, ruptures at the extensile end were related to cell length just as the rate of extension is related to cell length. The extension patterns of early log, mid-log, late log, and stationary phase cells from suspension cultures were found to approximate the linear growth patterns of Kubitschek and Clay (1986). The median length of cells was found to decline through the log phase in an unbalanced manner.     

Module-based construction of plasmids for chromosomal integration of the fission yeast Schizosaccharomyces pombe

Integration of an external gene into a fission yeast chromosome is useful to investigate the effect of the gene product. An easy way to knock-in a gene construct is use of an integration plasmid, which can be targeted and inserted to a chromosome through homologous recombination. Despite the advantage of integration, construction of integration plasmids is energy- and time-consuming, because there is no systematic library of integration plasmids with various promoters, fluorescent protein tags, terminators and selection markers; therefore, researchers are often forced to make appropriate ones through multiple rounds of cloning procedures. Here, we establish materials and methods to easily construct integration plasmids. We introduce a convenient cloning system based on Golden Gate DNA shuffling, which enables the connection of multiple DNA fragments at once: any kind of promoters and terminators, the gene of interest, in combination with any fluorescent protein tag genes and any selection markers. Each of those DNA fragments, called a ‘module’, can be tandemly ligated in the order we desire in a single reaction, which yields a circular plasmid in a one-step manner. The resulting plasmids can be integrated through standard methods for transformation. Thus, these materials and methods help easy construction of knock-in strains, and this will further increase the value of fission yeast as a model organism.     

Habitat quality,population dynamics,and group composition in Colobus Monkeys (Colobus guereza)

Data from an Ethiopian population of Colobus guerezashow that territory size is fixed by the high density of the population. Groups undergo fission when their size results in fewer than 10 trees per individual within the group’s territory. The daughter groups produced by fission emigrate into suboptimal habitat, which acts as a demographic sink. Comparative analyses using data from other East African populations demonstrate that mean territory size is inversely related to population density and that density, in turn, is a function of the size of the forest block. Since both group size and reproductive rates can be shown to be positively correlated with type of forest, it is concluded that this relationship reflects the fact that local population densities reach their ceiling more rapidly in small forest blocks because the animals’ access to alternative territories is limited. The number of males in a colobus group is shown to be a function of the number of females in it. Multimale groups have lower reproductive rates than one-male groups, probably because the stress generated by competition among the males causes temporary infertility among the females.     

“Wall-papering” and elaborate nest architecture in the ponerine antHarpegnathos saltator

Summary Excavation of 18 nests ofHarpegnathos saltator from southern India revealed an unusually complex architecture for a ponerine ant. The inhabited chambers are not deep in the ground. The uppermost chamber is protected by a thick vaulted roof, on the outside of which is an intervening space serving as isolation from the surrounding soil. In large colonies, the vaulted roof is extended into a shell which encloses several superimposed chambers. Little openings, which may be encircled by moulded flanges, occur in the upper region of the shell. The inside of the chambers is partly or completely lined with strips of empty cocoons. A refuse chamber is always found deeper than the inhabited chambers; live dipteran larvae (family Milichiidae) are typically present. These elaborate nests represent a large energetic investment, and we speculate therefore that nest emigration is unlikely in this species. Consequently, colony fission may never occur, unlike other ants where gamergates reproduce.     

Effect of papulacandin B on β-glucan synthesis in Schizosaccharomyces pombe

Abstract The antifungal antibiotic papulacandin β inhibited B(1,3)glucan-synthase activity, in vitro, from Schizosaccharomyces pombe . Levels of β(1,3)glucan-synthase from antibiotic-treated cultures were lower than the control cultures whereas mannan-synthase and β(1,3)glucanase activities were almost unaffected. The presence of an osmotic stabilizer reduced the inhibition of growth caused by the antibiotic. Addition of papulacandin β to a culture of S. pombe specifically inhibited incorporation of glucose into the β-glucan cell wall fraction. The fatty acids as well as the hydroxyl groups on the phenol residue of the papulacandin β molecule were essential for the inhibitory activity.     

Troop fission in free-ranging baboons in the Gombe Stream National Park, Tanzania.

During a 13 month field study of baboons in the Gombe Stream National Park, Tanzania, the troop under observation divided into two groups. One group left the main troop to become ultimately an independent troop with a separate range. The troop fission followed a period of instability in the male status hierarchy. The events prior to and during the troop fission are described in detail and the membership of the group which left is analyzed. Social bonds prior to the fission, including kinship, determined membership in the group which left. Aggression from members of the main troop played a major role in causing the group which left to establish a new range. This fission is compared with troop fissions observed in rhesus and Japances macaques.