mini spindles: A Gene Encoding a Conserved Microtubule-Associated Protein Required for the Integrity of the Mitotic Spindle in Drosophila

We describe a new Drosophila gene, mini spindles (msps) identified in a cytological screen for mitotic mutant. Mutation in msps disrupts the structural integrity of the mitotic spindle, resulting in the formation of one or more small additional spindles in diploid cells. Nucleation of microtubules from centrosomes, metaphase alignment of chromosomes, or the focusing of spindle poles appears much less affected. The msps gene encodes a 227-kD protein with high similarity to the vertebrate microtubule-associated proteins (MAPs), human TOGp and Xenopus XMAP215, and with limited similarity to the Dis1 and STU2 proteins from fission yeast and budding yeast. Consistent with their sequence similarity, Msps protein also associates with microtubules in vitro. In the embryonic division cycles, Msps protein localizes to centrosomal regions at all mitotic stages, and spreads over the spindles during metaphase and anaphase. The absence of centrosomal staining in interphase of the cellularized embryos suggests that the interactions between Msps protein and microtubules or centrosomes may be regulated during the cell cycle

Geostatistical analysis of mesoscale spatial variability and error in SeaWiFS and MODIS/Aqua global ocean color data

© The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Journal of Geophysical Research: Oceans 123 (2018): 22–39, doi:10.1002/2017JC013023.Mesoscale (10–300 km, weeks to months) physical variability strongly modulates the structure and dynamics of planktonic marine ecosystems via both turbulent advection and environmental impacts upon biological rates. Using structure function analysis (geostatistics), we quantify the mesoscale biological signals within global 13 year SeaWiFS (1998–2010) and 8 year MODIS/Aqua (2003–2010) chlorophyll a ocean color data (Level-3, 9 km resolution). We present geographical distributions, seasonality, and interannual variability of key geostatistical parameters: unresolved variability or noise, resolved variability, and spatial range. Resolved variability is nearly identical for both instruments, indicating that geostatistical techniques isolate a robust measure of biophysical mesoscale variability largely independent of measurement platform. In contrast, unresolved variability in MODIS/Aqua is substantially lower than in SeaWiFS, especially in oligotrophic waters where previous analysis identified a problem for the SeaWiFS instrument likely due to sensor noise characteristics. Both records exhibit a statistically significant relationship between resolved mesoscale variability and the low-pass filtered chlorophyll field horizontal gradient magnitude, consistent with physical stirring acting on large-scale gradient as an important factor supporting observed mesoscale variability. Comparable horizontal length scales for variability are found from tracer-based scaling arguments and geostatistical decorrelation. Regional variations between these length scales may reflect scale dependence of biological mechanisms that also create variability directly at the mesoscale, for example, enhanced net phytoplankton growth in coastal and frontal upwelling and convective mixing regions. Global estimates of mesoscale biophysical variability provide an improved basis for evaluating higher resolution, coupled ecosystem-ocean general circulation models, and data assimilation.NASA's Ocean Biology and Biogeochemistry Grant Numbers: NNG05GG30G, NNG05GR34G, NNX14AM36G, NNX14AL86G, NNX15AE65G; Ocean Biology Processing Group (OBPG) at NASA's Goddard Space Flight Cente

Antigenic variation in African trypanosomes: the importance of chromosomal and nuclear context in VSG expression control.

African trypanosomes are lethal human and animal parasites that use antigenic variation for evasion of host adaptive immunity. To facilitate antigenic variation, trypanosomes dedicate approximately one third of their nuclear genome, including many minichromosomes, and possibly all sub-telomeres, to variant surface glycoprotein (VSG) genes and associated sequences. Antigenic variation requires transcription of a single VSG by RNA polymerase I (Pol-I), with silencing of other VSGs, and periodic switching of the expressed gene, typically via DNA recombination with duplicative translocation of a new VSG to the active site. Thus, telomeric location, epigenetic controls and monoallelic transcription by Pol-I at an extranucleolar site are prominent features of VSGs and their expression, with telomeres, chromatin structure and nuclear organization all making vitally important contributions to monoallelic VSG expression control and switching. We discuss VSG transcription, recombination and replication control within this chromosomal and sub-nuclear context

Mutations in the Drosophila melanogaster gene three rows permit aspects of mitosis to continue in the absence of chromatid segregation

We have cloned the three rows (thr) gene, by a combination of chromosome microdissection and P element tagging. We describe phenotypes of embryos homozygous for mutations at the thr locus. Maternal mRNA and protein appear to be sufficient to allow 14 rounds of mitosis in embryos homozygous for thr mutations. However, a small percentage of cells in syncytial blastoderm stage thr embryos sink into the interior of the embryo as if they have failed to divide properly. Following cellularisation all cells complete mitosis 14 normally. All cells become delayed at mitosis 15 with their chromosomes remaining aligned on the spindle in a metaphase-like configuration, even though both cyclins A and B have both been degraded. As cyclin B degradation occurs at the metaphase-anaphase transition, subsequent to the microtubule integrity checkpoint, the delay induced by mutations at the thr locus defines a later point in mitotic progression. Chromosomes in the cells of thr embryos do not undertake anaphase separation, but remain at the metaphase plate. Subsequently they decondense. A subset of nuclei go on to replicate their DNA but there is no further mitotic division

Mutations in Drosophila Greatwall/Scant Reveal Its Roles in Mitosis and Meiosis and Interdependence with Polo Kinase

Polo is a conserved kinase that coordinates many events of mitosis and meiosis, but how it is regulated remains unclear. Drosophila females having only one wild-type allele of the polo kinase gene and the dominant Scant mutation produce embryos in which one of the centrosomes detaches from the nuclear envelope in late prophase. We show that Scant creates a hyperactive form of Greatwall (Gwl) with altered specificity in vitro, another protein kinase recently implicated in mitotic entry in Drosophila and Xenopus. Excess Gwl activity in embryos causes developmental failure that can be rescued by increasing maternal Polo dosage, indicating that coordination between the two mitotic kinases is crucial for mitotic progression. Revertant alleles of Scant that restore fertility to polo–Scant heterozygous females are recessive alleles or deficiencies of gwl; they show chromatin condensation defects and anaphase bridges in larval neuroblasts. One recessive mutant allele specifically disrupts a Gwl isoform strongly expressed during vitellogenesis. Females hemizygous for this allele are sterile, and their oocytes fail to arrest in metaphase I of meiosis; both homologues and sister chromatids separate on elongated meiotic spindles with little or no segregation. This allelic series of gwl mutants highlights the multiple roles of Gwl in both mitotic and meiotic progression. Our results indicate that Gwl activity antagonizes Polo and thus identify an important regulatory interaction of the cell cycle

Assessing the skill of a high-resolution marine biophysical model using geostatistical analysis of mesoscale ocean chlorophyll variability from field observations and remote sensing

© The Author(s), 2021. This article is distributed under the terms of the Creaive Commons Attribution License. The definitive version was published in Eveleth, R., Glover, D. M., Long, M. C., Lima, I. D., Chase, A. P., & Doney, S. C. . Assessing the skill of a high-resolution marine biophysical model using geostatistical analysis of mesoscale ocean chlorophyll variability from field observations and remote sensing. Frontiers in Marine Science, 8, (2021): 612764, https://doi.org/10.3389/fmars.2021.612764.High-resolution ocean biophysical models are now routinely being conducted at basin and global-scale, opening opportunities to deepen our understanding of the mechanistic coupling of physical and biological processes at the mesoscale. Prior to using these models to test scientific questions, we need to assess their skill. While progress has been made in validating the mean field, little work has been done to evaluate skill of the simulated mesoscale variability. Here we use geostatistical 2-D variograms to quantify the magnitude and spatial scale of chlorophyll a patchiness in a 1/10th-degree eddy-resolving coupled Community Earth System Model simulation. We compare results from satellite remote sensing and ship underway observations in the North Atlantic Ocean, where there is a large seasonal phytoplankton bloom. The coefficients of variation, i.e., the arithmetic standard deviation divided by the mean, from the two observational data sets are approximately invariant across a large range of mean chlorophyll a values from oligotrophic and winter to subpolar bloom conditions. This relationship between the chlorophyll a mesoscale variability and the mean field appears to reflect an emergent property of marine biophysics, and the high-resolution simulation does poorly in capturing this skill metric, with the model underestimating observed variability under low chlorophyll a conditions such as in the subtropics.This work was supported in part by the National Aeronautics and Space Administration (NASA) as part of the North Atlantic Aerosol and Marine Ecosystems Study (NAAMES; NASA grant 80NSSC18K0018). The CESM project is supported by the National Science Foundation and the Office of Science (BER) of the United States Department of Energy. Computing resources were provided by the Climate Simulation Laboratory at NCAR’s Computational and Information Systems Laboratory (CISL), sponsored by the National Science Foundation and other agencies. This research was enabled by CISL compute and storage resources

Low-resolution genome map of the malaria mosquito Anopheles gambiae

We have microdissected divisions of the Anopheles gambiae polytene chromosomes, digested the DNAs with a restriction enzyme, and PCR-amplified the DNA fragments to generate a set of pooled probes, each corresponding to approximately 2% of the mosquito genome. These divisional probes were shown to have high complexity. Except for those derived from near the centromeres, they hybridize specifically with their chromosomal sites of origin. Thus, they can be used to map cloned DNAs by a dot blot procedure, which is much more convenient than in situ hybridization to polytene chromosomes. We discuss additional potential uses of these probes, such as easier isolation of molecular markers and genes, including those that cross-hybridize with clones available from other insects. It is expected that the probes will substantially accelerate molecular genetic analysis of this most important malaria vector

Low-resolution genome map of the malaria mosquito Anopheles gambiae

We have microdissected divisions of the Anopheles gambiae polytene chromosomes, digested the DNAs with a restriction enzyme, and PCR-amplified the DNA fragments to generate a set of pooled probes, each corresponding to approximately 2% of the mosquito genome. These divisional probes were shown to have high complexity. Except for those derived from near the centromeres, they hybridize specifically with their chromosomal sites of origin. Thus, they can be used to map cloned DNAs by a dot blot procedure, which is much more convenient than in situ hybridization to polytene chromosomes. We discuss additional potential uses of these probes, such as easier isolation of molecular markers and genes, including those that cross-hybridize with clones available from other insects. It is expected that the probes will substantially accelerate molecular genetic analysis of this most important malaria vector