Cnd2 has dual roles in mitotic condensation and interphase

Chromosome condensation requires condensin, which comprises five subunits. Two of these subunits--both being structural maintenance of chromosome (SMC) proteins-are coiled-coils with globular terminal domains that interact with ATP and DNA. The remaining three, non-SMC subunits also have essential, albeit undefined, roles in condensation. Here we report that Cnd2 (ref. 6), a non-SMC subunit of fission yeast similar to Drosophila Barren and the budding yeast protein Brn1 (refs 8, 9), is required for both interphase and mitotic condensation. In cnd2-1 mutants, ultraviolet-induced DNA damage is not repaired, and cells arrested by hydroxyurea do not recover. A definitive defect of interphase is abolishment of Cds1 (a checkpoint kinase) activation in the presence of hydroxyurea in both cnd2-1 mutant cells and in cells where other condensin subunits have been genetically disrupted. In the absence of hydroxyurea, a G2 checkpoint delay occurred in cnd2-1 mutants in a manner dependent on Cds1 and ATM-like Rad3, but not Chk1 (refs 10-13), before the mitotic condensation defect. Furthermore, cnd2-1 was synthetic-lethal with mutations of excision repair, RecQ helicase and DNA replication enzymes. These interphase and mitotic defects provide insight into the mechanistic role of non-SMC subunits that interact with the globular SMC domains in the heteropentameric holocomplex.     

Effect of charge on growth of droplets with ionic condensation nuclei

A growth kinetics model of droplets with ionic condensation nuclei based on the chemical potential and the kinetic equation of mass transfer is established. The model is simplified and the effect of charge on droplet growth is examined. The theoretical results show that the critical radius for droplet growth with an ionic condensation nucleus is less than that of those without an ionic nucleus. Furthermore, our results also indicate that if the initial droplet with an ionic nucleus has a radius shorter than the critical radius, the droplet will not vanish, but will reach a steady-state radius. As the ionic charge increases, the critical radius for droplet growth will decrease and the corresponding steady-state droplet radius will increase. In addition we show that once a critical charge is reached, all droplets will grow regardless of initial radius.     

Intranuclear injection of anti-actin antibodies into Xenopus oocytes blocks chromosome condensation

The role of contractile proteins in the structural organisation of the interphase nucleus and of metaphase chromosomes is largely unknown. Actin has been found in interphase nuclei of different species, especially in association with condensed chromatin. In the germinal vesicle (nucleus) of Xenopus oocytes, actin has been localised in the nuclear gel supporting the chromosomes and the extrachromosomal nucleoli. It has been reported that the premeiotic lampbrush chromosomes in these germinal vesicles are positively stained for actin and tubulin by the immunoperoxidase technique. Moreover, the longitudinal contraction of these chromosomes is ATP dependent. Therefore it has been suggested that actin participates in the structural organisation of the highly specialised lampbrush chromosomes. However, actin is not a major component of the metaphase chromosome scaffold. The results reported here suggest that actin is involved in the condensation of Xenopus chromosomes.     

MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells

The mitotic checkpoint protein hsMad2 is required to arrest cells in mitosis when chromosomes are unattached to the mitotic spindle. The presence of a single, lagging chromosome is sufficient to activate the checkpoint, producing a delay at the metaphase-anaphase transition until the last spindle attachment is made. Complete loss of the mitotic checkpoint results in embryonic lethality owing to chromosome mis-segregation in various organisms. Whether partial loss of checkpoint control leads to more subtle rates of chromosome instability compatible with cell viability remains unknown. Here we report that deletion of one MAD2 allele results in a defective mitotic checkpoint in both human cancer cells and murine primary embryonic fibroblasts. Checkpoint-defective cells show premature sister-chromatid separation in the presence of spindle inhibitors and an elevated rate of chromosome mis-segregation events in the absence of these agents. Furthermore, Mad2+/- mice develop lung tumours at high rates after long latencies, implicating defects in the mitotic checkpoint in tumorigenesis.     

Focal points for chromosome condensation and decondensation revealed by three-dimensional in vivo time-lapse microscopy

Although the dynamic behaviour of chromosomes has been extensively studied in their condensed state during mitosis, chromosome behaviour during the transition to and from interphase has not been well documented. Previous electron microscopic studies suggest that chromosomes condense in a non-uniform fashion at the nuclear periphery. But chromosome condensation is a complicated and dynamic process and requires continuous observation in living tissues to be fully understood. Using a recently developed three-dimensional time-lapse fluorescence microscopy technique, we have observed chromosomes as they relax from telophase, through interphase, until their condensation at the next prophase. This technique has been improved to produce higher-resolution images by implementing new stereographic projection and computational processing protocols. These studies have revealed that chromosomal regions on the nuclear envelope, distinct from the centromeres and telomeres, serve as foci for the decondensation and condensation of diploid chromosomes. The relative positions of the late decondensation sites at the beginning of interphase appear to correspond to the early condensation sites at the subsequent prophase.     

Absence of oestradiol concentration in cell nuclei of LHRH-immunoreactive neurones

Oestrogen, acting in both the brain and pituitary, has a critical role in regulating the reproductive cycle in most mammals. In the brain, oestrogen regulates the release of luteinizing hormone-releasing hormone (LHRH) partly through a mechanism that is blocked by inhibitors of DNA-dependent RNA synthesis or protein synthesis. The distributions of oestrogen-concentrating neurones and of LHRH neurones overlap. The present study was undertaken to determine whether genomic effects of oestrogen mediated by nuclear oestradiol concentration include a direct effect on LHRH-containing neurones. During extensive studies in which the immunocytochemical method for localizing LHRH neurones was optimized and made compatible with the autoradiographic method for detecting oestrogen-concentrating neurones, doubly-labelled cells were very rarely seen. This suggests that genomic regulatory effects of oestrogen which depend on nuclear retention are not exerted directly on most LHRH neurones, but rather must be mediated by another class of neurones.