Integrin trafficking regulated by Rab21 is necessary for cytokinesis

Adherent cells undergo remarkable changes in shape during cell division. However, the functional interplay between cell adhesion turnover and the mitotic machinery is poorly understood. The endo/exocytic trafficking of integrins is regulated by the small GTPase Rab21, which associates with several integrin alpha subunits. Here, we show that targeted trafficking of integrins to and from the cleavage furrow is required for successful cytokinesis, and that this is regulated by Rab21. Rab21 activity, integrin-Rab21 association, and integrin endocytosis are all necessary for normal cytokinesis, which becomes impaired when integrin-mediated adhesion at the cleavage furrow fails. We also describe a chromosomal deletion and loss of Rab21 gene expression in human cancer, which leads to the accumulation of multinucleate cells. Importantly, reintroduction of Rab21 rescued this phenotype. In conclusion, Rab21-regulated integrin trafficking is essential for normal cell division, and its defects may contribute to multinucleation and genomic instability, which are hallmarks of cancer.     

The effect of Chinese tallow tree (Sapium sebiferum) ecotype on soil–plant system carbon and nitrogen processes

The EICA hypothesis predicts that shifts in allocation of invasive plants give rise to higher growth rates and lower herbivore defense levels in their introduced range than conspecifics in their native range. These changes in traits of invasive plants may also affect ecosystem processes. We conducted an outdoor pot experiment with Chinese tallow tree (Sapium sebiferum, Euphorbiaceae) seedlings from its native (Jiangsu, China, native ecotype) and introduced ranges (Texas, USA, invasive ecotype) to compare their relative performances in its native range and to examine ecotype effects on soil processes with and without fertilization. Consistent with predictions, plant (shoot and root) mass was significantly greater and leaf defoliation tended to be higher, while the root:shoot ratio was lower for the invasive ecotype relative to the native ecotype. Seasonal amounts of soil–plant system CO₂ and N₂O emissions were higher for the invasive ecotype than for the native ecotype. Soil respiration rates and N₂O emission increases from fertilization were also greater for the invasive ecotype than for the native ecotype, while shoot-specific respiration rates (g CO₂–C g⁻¹ C day⁻¹) did not differ between ecotypes. Further, soil inorganic N (ammonium and nitrate) was higher, but soil total N was lower for soils with the invasive ecotype than soils with the native ecotype. Compared with native ecotypes, therefore, invasive ecotypes may have developed a competition advantage in accelerating soil processes and promoting more nitrogen uptake through soil–plant direct interaction. The results of this study suggest that soil and ecosystem processes accelerated by variation in traits of invasive plants may have implications for their invasiveness.     

Aging bone marrow mesenchymal stromal cells have altered membrane glycerophospholipid composition and functionality

Human mesenchymal stem/stromal cells (hMSC) are increasingly used in advanced cellular therapies. The clinical use of hMSCs demands sequential cell expansions. As it is well established that membrane glycerophospholipids (GPL) provide precursors for signaling lipids that modulate cellular functions, we studied the effect of the donor''s age and cell doublings on the GPL profile of human bone marrow MSC (hBMSC). The hBMSCs, which were harvested from five young and five old adults, showed clear compositional changes during expansion seen at the level of lipid classes, lipid species, and acyl chains. The ratio of phosphatidylinositol to phosphatidylserine increased toward the late-passage samples. Furthermore, 20:4n-6-containing species of phosphatidylcholine and phosphatidylethanolamine accumulated while the species containing monounsaturated fatty acids (FA) decreased during passaging. Additionally, in the total FA pool of the cells, 20:4n-6 increased, which happened at the expense of n-3 polyunsaturated FAs, especially 22:6n-3. The GPL and FA correlated with the decreased immunosuppressive capacity of hBMSCs during expansion. Our observations were further supported by alterations in the gene expression levels of several enzymes involved in lipid metabolism and immunomodulation. The results show that extensive expansion of hBMSCs harmfully modulates membrane GPLs, affecting lipid signaling and eventually impairing functionality.     

Somatic progenitor cell vulnerability to mitochondrial DNA mutagenesis underlies progeroid phenotypes in Polg mutator mice

Somatic stem cell (SSC) dysfunction is typical for different progeroid phenotypes in mice with genomic DNA repair defects. MtDNA mutagenesis in mice with defective Polg exonuclease activity also leads to progeroid symptoms, by an unknown mechanism. We found that Polg-Mutator mice had neural (NSC) and hematopoietic progenitor (HPC) dysfunction already from embryogenesis. NSC self-renewal was decreased in vitro, and quiescent NSC amounts were reduced in vivo. HPCs showed abnormal lineage differentiation leading to anemia and lymphopenia. N-acetyl-L-cysteine treatment rescued both NSC and HPC abnormalities, suggesting that subtle ROS/redox changes, induced by mtDNA mutagenesis, modulate SSC function. Our results show that mtDNA mutagenesis affected SSC function early but manifested as respiratory chain deficiency in nondividing tissues in old age. Deletor mice, having mtDNA deletions in postmitotic cells and no progeria, had normal SSCs. We propose that SSC compartment is sensitive to mtDNA mutagenesis, and that mitochondrial dysfunction in SSCs can underlie progeroid manifestations.     

Structure and dynamics of a human myelin protein P2 portal region mutant indicate opening of the β barrel in fatty acid binding proteins

Background

Myelin is a multilayered proteolipid sheath wrapped around selected axons in the nervous system. Its constituent proteins play major roles in forming of the highly regular membrane structure. P2 is a myelin-specific protein of the fatty acid binding protein (FABP) superfamily, which is able to stack lipid bilayers together, and it is a target for mutations in the human inherited neuropathy Charcot-Marie-Tooth disease. A conserved residue that has been proposed to participate in membrane and fatty acid binding and conformational changes in FABPs is Phe57. This residue is thought to be a gatekeeper for the opening of the portal region upon ligand entry and egress.

Results

We performed a structural characterization of the F57A mutant of human P2. The mutant protein was crystallized in three crystal forms, all of which showed changes in the portal region and helix α2. In addition, the behaviour of the mutant protein upon lipid bilayer binding suggested more unfolding than previously observed for wild-type P2. On the other hand, membrane binding rendered F57A heat-stable, similarly to wild-type P2. Atomistic molecular dynamics simulations showed opening of the side of the discontinuous β barrel, giving important indications on the mechanism of portal region opening and ligand entry into FABPs. The results suggest a central role for Phe57 in regulating the opening of the portal region in human P2 and other FABPs, and the F57A mutation disturbs dynamic cross-correlation networks in the portal region of P2.

Conclusions

Overall, the F57A variant presents similar properties to the P2 patient mutations recently linked to Charcot-Marie-Tooth disease. Our results identify Phe57 as a residue regulating conformational changes that may accompany membrane surface binding and ligand exchange in P2 and other FABPs.

     

Population Dynamics and Potential for Biological Control of an Exotic Invasive Shrub in Hawaiian Rainforests

Introduction of biological control agents to reduce the abundance of exotic invasive plant species is often considered necessary but risky. I used matrix projection models to investigate the current population dynamics of Clidemia hirta (Melastomataceae), an invasive shrub, in two rainforest stands on the island of Hawaii and to predict the efficacy of hypothetical biological control agents in reducing population growth rates. Stage-structured matrix models were parameterized with field data collected over 3 years from 2906 C. hirta plants in a recently invaded forest with an open overstory (Laupahoehoe) and 600 plants in a less recently invaded forest with a closed canopy (Waiakea). Asymptotic population growth rates (λ) for both populations in all years were greater than one, demonstrating that both populations were growing. Composite elasticities were high for the seedling life-history stage and fecundity, and near-term demographic elasticities suggested that changes in seedling survival would have the largest effect on population size in the short term. However, simulations showed that almost 100% of seedlings or new recruits produced per reproductive adult would have to be destroyed to cause populations to go locally extinct under current environmental conditions. Herbivores or pathogens that decrease survival across all vegetative stages by 12% at Waiakea and 64% at Laupahoehoe were projected to cause the populations to decline. Thus, biocontrol agents that reduce survival of multiple life-history stages rather than seed production should be pursued to control C. hirta in Hawaiian rainforests.     

Classification of European Mtdnas from an Analysis of Three European Populations

Mitochondrial DNA (mtDNA) sequence variation was examined in Finns, Swedes and Tuscans by PCR amplification and restriction analysis. About 99% of the mtDNAs were subsumed within 10 mtDNA haplogroups (H, I, J, K, M, T, U, V, W, and X) suggesting that the identified haplogroups could encompass virtually all European mtDNAs. Because both hypervariable segments of the mtDNA control region were previously sequenced in the Tuscan samples, the mtDNA haplogroups and control region sequences could be compared. Using a combination of haplogroup-specific restriction site changes and control region nucleotide substitutions, the distribution of the haplogroups was surveyed through the published restriction site polymorphism and control region sequence data of Caucasoids. This supported the conclusion that most haplogroups observed in Europe are Caucasoid-specific, and that at least some of them occur at varying frequencies in different Caucasoid populations. The classification of almost all European mtDNA variation in a number of well defined haplogroups could provide additional insights about the origin and relationships of Caucasoid populations and the process of human colonization of Europe, and is valuable for the definition of the role played by mtDNA backgrounds in the expression of pathological mtDNA mutations