Dynamic regulation of GEF-H1 localization at microtubules by Par1b/MARK2

Par1b/MARK2 is a serine/threonine kinase that plays key roles in the development of cell polarity, but its precise mechanism of action remains unknown. Here we report that GEF-H1, a guanine nucleotide exchange factor for Rho-family small GTPases, is a novel substrate for Par1b. GEF-H1 directly associates with microtubules via its N-terminal C1 domain, which is known to regulate the activity of GEF-H1. Ectopically expressed GEF-H1 markedly promotes stabilization of microtubules, resulting in acetylation of microtubules. We find that Par1b phosphorylates GEF-H1 at three serine residues conserved in vertebrates and releases GEF-H1 from microtubules, which abrogates stabilization and acetylation of microtubules induced by GEF-H1 overexpression. The alanine mutant for the three phosphorylation sites (3SA) of GEF-H1 strongly induces stabilization and acetylation of microtubules, which was resistant to Par1b. Time-lapse imaging analyses reveal that GFP-fused GEF-H1 dynamically moved on microtubules from one protrusion to another, whereas the 3SA mutant was static. These data suggest that Par1b-phosphorylation regulates turnover of GEF-H1 localization by regulating its interaction with microtubules, which may contribute to cell polarization.     

Independent evolution of a new allele of F<Subscript>1</Subscript> pollen sterility gene <Emphasis Type="Italic">S27</Emphasis> encoding mitochondrial ribosomal protein L27 in <Emphasis Type="Italic">Oryza nivara</Emphasis>

Loss of function of duplicated genes plays an important role in the evolution of postzygotic reproductive isolation. The widespread occurrence of gene duplication followed by rapid loss of function of some of the duplicate gene copies suggests the independent evolution of loss-of-function alleles of duplicate genes in divergent lineages of speciation. Here, we found a novel loss-of-function allele of S27 in the Asian annual wild species Oryza nivara, designated S27-niv ^s , that leads to F₁ pollen sterility in a cross between O. sativa and O. nivara. Genetic linkage analysis and complementation analysis demonstrated that S27-niv ^s lies at the same locus as the previously identified S27 locus and S27-niv ^s is a loss-of-function allele of S27. S27-niv ^s is composed of two tandem mitochondrial ribosomal protein L27 genes (mtRPL27a and mtRPL27b), both of which are inactive. The coding and promoter regions of S27-niv ^s showed a number of nucleotide differences from the functional S27-T65 ⁺ allele. The structure of S27-niv ^s is different from that of a previously identified null S27 allele, S27-glum ^s , in the South American wild rice species O. glumaepatula, in which mtRPL27a and mtRPL27b are absent. These results show that the mechanisms for loss-of-function of S27-niv ^s and S27-glum ^s are different. Our results provide experimental evidence that different types of loss-of-function alleles are distributed in geographically and phylogenetically isolated species and represent a potential mechanism for postzygotic isolation in divergent species.     

Unmasking of LPA1 receptor-mediated migration response to lysophosphatidic acid by interleukin-1β-induced attenuation of Rho signaling pathways in rat astrocytes

Action mechanism of lipopolysaccharide (LPS), interleukin-1β (IL-1β), and lysophosphatidic acid (LPA) to regulate motility, an important process of astrogliosis, was investigated in rat astrocytes. While LPA exerted no significant effect on the cell migration, the prior treatment of the cells with LPS or IL-1β resulted in the appearance of migration activity in response to LPA. The LPS induction of the migration response to LPA was associated with the production of IL-1β precursor protein and inhibited by the IL-1 receptor antagonist. The IL-1β treatment also allowed LPA to activate Rac1. The LPA-induced Rac1 activation and migration were inhibited by pertussis toxin, a small interfering RNA specific to LPA(1) receptors, and LPA(1) receptor antagonists, including Ki16425. However, the IL-1β treatment had no appreciable effect on LPA(1) receptor mRNA expression and LPA-induced activation of ERK, Akt, and proliferation. The induction of the migration response to LPA by IL-1β was inhibited by a constitutively active RhoA. Moreover, LPA significantly activated RhoA through the LPA(1) receptor in the control cells but not in the IL-1β-treated cells. These results suggest that IL-1β inhibits the LPA(1) receptor-mediated Rho signaling through the IL-1 receptor, thereby disclosing the LPA(1) receptor-mediated G(i) protein/Rac/migration pathway.     

The potential biodiversity of Ayu,as evidenced by differences in its early development and growth between Vietnam and Japan

To examine regional specific diversity in development and growth of Ayu (Plecoglossus altivelis) larvae, we collected and compared collections from the Kalong estuary in Vietnam, and the Shimanto and Muko estuary, and the Niigata coast in Japan. Among the four areas, most of the morphometrics through ontogeny were similar except that the snout tended to be shorter and the anus hardly migrated in Kalong larvae. The snout length increased gradually with growth in the Vietnamese larvae, while this value increased significantly until ca. 10 mm BL, subsequently being constant up to 30 mm body length (BL) in the Japanese larvae. The water temperature when the larvae were collected was higher in the Kalong than in most of the Japan sites. Growth-rates estimated from otolith increments were from highest to lowest, Niigata (mean?=?0.54 mm/day), Kalong (0.47), Shimanto (0.38) and Muko (0.34). The higher growth-rates were obtained not in Niigata of highest latitudinal region, but in Kalong of lowest latitudinal region. This indicates that Ayu could experience their early developmental stages from the cool temperate to tropical regions, implying the potential biodiversity of this fish species in the world.     

Two intromittent organs in Zorotypus caudelli (Insecta,Zoraptera): the paradoxical coexistence of an extremely long tube and a large spermatophore

Very unusual genitalia of the species Zorotypus caudelli are described. It contains the unique configuration of two different intromittent organs, one of them strongly elongated. Hyper elongated genitalia are known in different groups of insects. Males have to accommodate these unwieldy structures in the limited spaces of the abdomen and manipulate them acutely during copulation. A crucial question is how do species with elongated genitalia cope with these requirements? To investigate this, we studied key features enabling storage, insertion, and withdrawal of the elongated genitalia. The co‐existence of an elongated narrow tube and a bulky spermatophore is a highly unusual and apparently paradoxical condition. However, we demonstrate that the tube is not involved in sperm transmission, whereas the large spermatophore is transferred to females by a membranous fold of the genitalia. The movement of the spermatophore is caused by haemolymph pressure, which likely also promotes the insertion of both intromittent organs. A comparison with the genital anatomy and reproductive mode in related groups suggests that the elongated tube and its accommodating pouch is a de novo structure, and that the ancestral sperm transport via spermatophore is a preadaptive condition for the acquisition of this unusual structure. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 40–54.     

Contribution of Neuropilin-1 in Radiation-Survived Subclones of NSCLC Cell Line H1299

Non-small cell lung cancer (NSCLC) is an aggressive lung cancer accounting for approximately 85% of all lung cancer patients. For the patients with Stages IIIA, IIIB, and IIIC, the 5-year survival is low though with the combination with radiotherapy and chemotherapy. In addition, the occurrence of tumor cells (repopulated tumors) that survive irradiation remains a challenge. In our previous report, we subcloned the radiation-surviving tumor cells (IR cells) using the human NSCLC cell line, H1299, and found that the expression of neuropilin-1 (NRP-1) was upregulated in IR cells by the microarray analysis. Here, we investigated the contribution of neuropilin-1 to changes in the characteristics of IR cells. Although there were no differences in angiogenic activity in the tube formation assay between parental and IR cells, the cell motility was increased in IR cells compared to parental cells in the cell migration assay. This enhanced cell motility was suppressed by pretreatment with anti-NRP-1 antibody. Although further studies are necessary to identify other molecules associated with NRP-1, the increase in cellular motility in IR cells might be due to the contribution of NRP-1. Inhibition of NRP-1 would help control tumor malignancy in radiation-surviving NSCLC.     

Seasonal changes in the deleterious effects of solar ultraviolet-B radiation on eggs of the twospotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae)

Solar ultraviolet-B (UVB) radiation has deleterious effects on plant-dwelling mites. We assessed the biological effects of UVB radiation on the eggs of the twospotted spider mite, Tetranychus urticae Koch, under both near ambient (UV+) and UV-attenuated (UV−) conditions from spring to autumn and compared them to the effects of temperature and humidity. The ambient daily UVB irradiance increased from January to August and then decreased rapidly until December, whereas egg hatchability under UV+ was lowest in April (10.7%) and increased almost linearly until October (74.9–92.3%). In contrast, hatchability under UV− was consistently high (96.2–99.8%) through all seasons. For UV+, the stepwise multiple linear regression analysis supported the negative correlation of hatchability with cumulative UVB irradiance during egg periods (cumulative dose), but did not support that with the mean daily UVB irradiance (dose rate), suggesting that UVB-induced mortality in T. urticae eggs is cumulative dose dependent rather than dose rate dependent. The high mortality in April may have reflected the slower development caused by the relatively lower temperature and higher UVB radiation, increasing the cumulative dose, while the low mortality in October may have reflected the faster development caused by the relatively higher temperature and lower UVB radiation, decreasing the cumulative dose.