Identification and characterization of the ARP1 gene, a target for the human acute leukemia ALL1 gene

ALL1, the human homologue of Drosophila trithorax, is directly involved in human acute leukemias associated with abnormalities at 11q23. Using the differential display method, we isolated a gene that is down-regulated in All1 double-knockout mouse embryonic stem (ES) cells. The gene, designated ARP1 (also termed RIEG, Ptx2, or Otlx2), is a member of a family of homeotic genes containing a short motif shared with several homeobox genes. Using a bacterially synthesized All1 polypeptide encompassing the AT-hook motifs, we identified a 0.5-kb ARP1 DNA fragment that preferentially bound to the polypeptide. Within this DNA, a region of ≈100 bp was protected by the polypeptide from digestion with ExoIII and DNase I. Whole-mount in situ hybridization to early mouse embryos of 9.5–10.5 days indicated a complex pattern of Arp1 expression spatially overlapping with the expression of All1. Although the ARP1 gene is expressed strongly in bone marrow cells, no transcripts were detected in six leukemia cell lines with 11q23 translocations. These results suggest that ARP1 is up-regulated by the All1 protein, possibly through direct interaction with an upstream DNA sequence of the former. The results are also consistent with the suggestion that ALL1 chimeric proteins resulting from 11q23 abnormalities act in a dominant negative fashion.

High-Voltage Electron Tomography of Spindle Pole Bodies and Early Mitotic Spindles in the Yeast Saccharomyces cerevisiaeV⃞

The spindle pole body (SPB) is the major microtubule-organizing center of budding yeast and is the functional equivalent of the centrosome in higher eukaryotic cells. We used fast-frozen, freeze-substituted cells in conjunction with high-voltage electron tomography to study the fine structure of the SPB and the events of early spindle formation. Individual structures were imaged at 5–10 nm resolution in three dimensions, significantly better than can be achieved by serial section electron microscopy. The SPB is organized in distinct but coupled layers, two of which show ordered two-dimensional packing. The SPB central plaque is anchored in the nuclear envelope with hook-like structures. The minus ends of nuclear microtubules (MTs) are capped and are tethered to the SPB inner plaque, whereas the majority of MT plus ends show a distinct flaring. Unbudded cells containing a single SPB retain 16 MTs, enough to attach to each of the expected 16 chromosomes. Their median length is ∼150 nm. MTs growing from duplicated but not separated SPBs have a median length of ∼130 nm and interdigitate over the bridge that connects the SPBs. As a bipolar spindle is formed, the median MT length increases to ∼300 nm and then decreases to ∼30 nm in late anaphase. Three-dimensional models confirm that there is no conventional metaphase and that anaphase A occurs. These studies complement and extend what is known about the three-dimensional structure of the yeast mitotic spindle and further our understanding of the organization of the SPB in intact cells.

Defining the minimal length of sequence homology required for selective gene isolation by TAR cloning

The transformation-associated recombination (TAR) cloning technique allows selective and accurate isolation of chromosomal regions and genes from complex genomes. The technique is based on in vivo recombination between genomic DNA and a linearized vector containing homologous sequences, or hooks, to the gene of interest. The recombination occurs during transformation of yeast spheroplasts that results in the generation of a yeast artificial chromosome (YAC) containing the gene of interest. To further enhance and refine the TAR cloning technology, we determined the minimal size of a specific hook required for gene isolation utilizing the Tg.AC mouse transgene as a targeted region. For this purpose a set of vectors containing a B1 repeat hook and a Tg.AC-specific hook of variable sizes (from 20 to 800 bp) was constructed and checked for efficiency of transgene isolation by a radial TAR cloning. When vectors with a specific hook that was ≥60 bp were utilized, ∼2% of transformants contained circular YACs with the Tg.AC transgene sequences. Efficiency of cloning dramatically decreased when the TAR vector contained a hook of 40 bp or less. Thus, the minimal length of a unique sequence required for gene isolation by TAR is ∼60 bp. No transgene-positive YAC clones were detected when an ARS element was incorporated into a vector, demonstrating that the absence of a yeast origin of replication in a vector is a prerequisite for efficient gene isolation by TAR cloning.

NPH4, a Conditional Modulator of Auxin-Dependent Differential Growth Responses in Arabidopsis1

Although sessile in nature, plants are able to use a number of mechanisms to modify their morphology in response to changing environmental conditions. Differential growth is one such mechanism. Despite its importance in plant development, little is known about the molecular events regulating the establishment of differential growth. Here we report analyses of the nph4 (nonphototropic hypocotyl) mutants of Arabidopsis that suggest that the NPH4 protein plays a central role in the modulation of auxin-dependent differential growth. Results from physiological studies demonstrate that NPH4 activity is conditionally required for a number of differential growth responses, including phototropism, gravitropism, phytochrome-dependent hypocotyl curvature, apical hook maintenance, and abaxial/adaxial leaf-blade expansion. The nph4 mutants exhibited auxin resistance and severely impaired auxin-dependent gene expression, indicating that the defects associated with differential growth likely arise because of altered auxin responsiveness. Moreover, the auxin signaling events mediating phototropism are genetically correlated with the abundance of the NPH4 protein.

Jasmonic acid distribution and action in plants: regulation during development and response to biotic and abiotic stress.

Jasmonic acid (JA) is a naturally occurring growth regulator found in higher plants. Several physiological roles have been described for this compound (or a related compound, methyl jasmonate) during plant development and in response to biotic and abiotic stress. To accurately determine JA levels in plant tissue, we have synthesized JA containing 13C for use as an internal standard with an isotopic composition of [225]:[224] 0.98:0.02 compared with [225]:[224] 0.15:0.85 for natural material. GC analysis (flame ionization detection and MS) indicate that the internal standard is composed of 92% 2-(+/-)-[13C]JA and 8% 2-(+/-)-7-iso-[13C]JA. In soybean plants, JA levels were highest in young leaves, flowers, and fruit (highest in the pericarp). In soybean seeds and seedlings, JA levels were highest in the youngest organs including the hypocotyl hook, plumule, and 12-h axis. In soybean leaves that had been dehydrated to cause a 15% decrease in fresh weight, JA levels increased approximately 5-fold within 2 h and declined to approximately control levels by 4 h. In contrast, a lag time of 1-2 h occurred before abscisic acid accumulation reached a maximum. These results will be discussed in the context of multiple pathways for JA biosynthesis and the role of JA in plant development and responses to environmental signals.

New York Bay and Harbor, 1776 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Chart of the entrance of Hudson's River, from Sandy Hook to New York : with the banks, depths of water, sailing-marks, & ca. It was printed for Robt. Sayer & John Bennett, sea chart & map-sellers ... 1st June 1776. Scale [ca. 1:70,000]. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD83 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows coastal features such as lighthouses, rocks, channels, points, coves, islands, and more. Depths shown by soundings and shading. Relief shown by hachures. Includes notes. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New York Bay and Harbor, and vicinity, 1844 (Image 2 of 6) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of New-York Bay and Harbor and the environs : sheet no. 2, founded upon a trigonometrical survey under the direction of F.R. Hassler, superintendent of the Survey of the Coast of the United States ; triangulation by James Ferguson and Edmund Blunt, assistants ; the hydrography under the direction of Thomas R. Gedney, lieutenant U.S. Navy ; the topography by C. Renard and T.A. Jenkins assists. It was published by Survey of the Coast of the United States in 1844-1845. Scale 1:30,000. This layer is image 2 of 6 total images of the six sheet source map, representing the southeast portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD83 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, or other information associated with the principal map. This map shows coastal features such as lighthouses, buoys, beacons, rocks, channels, points, coves, islands, bottom soil types, wharves, and more. Includes also selected land features such as roads, drainage, land cover, forts, selected buildings, towns, and more. Relief shown by hachures. Depths are shown by soundings and shading. Includes text, table of currents and stations, notes, sailing directions, 4 coastal panoramas and 2 views of Sandy Hook Light. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New York Bay and Harbor, and vicinity, 1844 (Image 1 of 6) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of New-York Bay and Harbor and the environs : sheet no. 1, founded upon a trigonometrical survey under the direction of F.R. Hassler, superintendent of the Survey of the Coast of the United States ; triangulation by James Ferguson and Edmund Blunt, assistants ; the hydrography under the direction of Thomas R. Gedney, lieutenant U.S. Navy ; the topography by C. Renard and T.A. Jenkins assists. It was published by Survey of the Coast of the United States in 1844-1845. Scale 1:30,000. This layer is image 1 of 6 total images of the six sheet source map, representing the southwest portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD83 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, or other information associated with the principal map. This map shows coastal features such as lighthouses, buoys, beacons, rocks, channels, points, coves, islands, bottom soil types, wharves, and more. Includes also selected land features such as roads, drainage, land cover, forts, selected buildings, towns, and more. Relief shown by hachures. Depths are shown by soundings and shading. Includes text, table of currents and stations, notes, sailing directions, 4 coastal panoramas and 2 views of Sandy Hook Light. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New York Bay and Harbor, and vicinity, 1844 (Image 4 of 6) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of New-York Bay and Harbor and the environs : sheet no. 4, founded upon a trigonometrical survey under the direction of F.R. Hassler, superintendent of the Survey of the Coast of the United States ; triangulation by James Ferguson and Edmund Blunt, assistants ; the hydrography under the direction of Thomas R. Gedney, lieutenant U.S. Navy ; the topography by C. Renard and T.A. Jenkins assists. It was published by Survey of the Coast of the United States in 1844-1845. Scale 1:30,000. This layer is image 4 of 6 total images of the six sheet source map, representing the southwest portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD83 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, or other information associated with the principal map. This map shows coastal features such as lighthouses, buoys, beacons, rocks, channels, points, coves, islands, bottom soil types, wharves, and more. Includes also selected land features such as roads, drainage, land cover, forts, selected buildings, towns, and more. Relief shown by hachures. Depths are shown by soundings and shading. Includes text, table of currents and stations, notes, sailing directions, 4 coastal panoramas and 2 views of Sandy Hook Light. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New York Bay and Harbor, and vicinity, 1844 (Image 1 of 6) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of New-York Bay and Harbor and the environs : sheet no. 3, founded upon a trigonometrical survey under the direction of F.R. Hassler, superintendent of the Survey of the Coast of the United States ; triangulation by James Ferguson and Edmund Blunt, assistants ; the hydrography under the direction of Thomas R. Gedney, lieutenant U.S. Navy ; the topography by C. Renard and T.A. Jenkins assists. It was published by Survey of the Coast of the United States in 1844-1845. Scale 1:30,000. This layer is image 3 of 6 total images of the six sheet source map, representing the southwest portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD83 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, or other information associated with the principal map. This map shows coastal features such as lighthouses, buoys, beacons, rocks, channels, points, coves, islands, bottom soil types, wharves, and more. Includes also selected land features such as roads, drainage, land cover, forts, selected buildings, towns, and more. Relief shown by hachures. Depths are shown by soundings and shading. Includes text, table of currents and stations, notes, sailing directions, 4 coastal panoramas and 2 views of Sandy Hook Light. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New York Bay and Harbor, and vicinity, 1844 (Image 5 of 6) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of New-York Bay and Harbor and the environs : sheet no. 5, founded upon a trigonometrical survey under the direction of F.R. Hassler, superintendent of the Survey of the Coast of the United States ; triangulation by James Ferguson and Edmund Blunt, assistants ; the hydrography under the direction of Thomas R. Gedney, lieutenant U.S. Navy ; the topography by C. Renard and T.A. Jenkins assists. It was published by Survey of the Coast of the United States in 1844-1845. Scale 1:30,000. This layer is image 5 of 6 total images of the six sheet source map, representing the southwest portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD83 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, or other information associated with the principal map. This map shows coastal features such as lighthouses, buoys, beacons, rocks, channels, points, coves, islands, bottom soil types, wharves, and more. Includes also selected land features such as roads, drainage, land cover, forts, selected buildings, towns, and more. Relief shown by hachures. Depths are shown by soundings and shading. Includes text, table of currents and stations, notes, sailing directions, 4 coastal panoramas and 2 views of Sandy Hook Light. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New York Bay and Harbor, and vicinity, 1844 (Image 6 of 6) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of New-York Bay and Harbor and the environs : sheet no. 6, founded upon a trigonometrical survey under the direction of F.R. Hassler, superintendent of the Survey of the Coast of the United States ; triangulation by James Ferguson and Edmund Blunt, assistants ; the hydrography under the direction of Thomas R. Gedney, lieutenant U.S. Navy ; the topography by C. Renard and T.A. Jenkins assists. It was published by Survey of the Coast of the United States in 1844-1845. Scale 1:30,000. This layer is image 6 of 6 total images of the six sheet source map, representing the southwest portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD83 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, or other information associated with the principal map. This map shows coastal features such as lighthouses, buoys, beacons, rocks, channels, points, coves, islands, bottom soil types, wharves, and more. Includes also selected land features such as roads, drainage, land cover, forts, selected buildings, towns, and more. Relief shown by hachures. Depths are shown by soundings and shading. Includes text, table of currents and stations, notes, sailing directions, 4 coastal panoramas and 2 views of Sandy Hook Light. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

Philadelphia, Pennsylvania (West), and vicinity, 1956 (Raster Image) (Image 2 of 2)

This layer is a georeferenced raster image of the United States Geological Survey sheet map set entitled: Philadelphia and vicinity, east, 1955 (and west, 1956) (Pennsylvania - New Jersey) by the Geological Survey. It was published in 1958. Scale 1:24,000. Covers Philadelphia and portions of adjacent counties. Mapped by the Geological Survey and U.S. Coast and Geodetic Survey. Compiled from 1:24,000 scale maps of Ambler 1952, Lansdale 1951, Collegeville 1951, Valley Forge 1952, Norristown 1952, Germantown 1952, Philadelphia 1949, Lansdowne 1956, Media 1955, Marcus Hook 1953, Bridgeport 1953, and Woodbury 1949 1953 7.5 minute quadrangles. This layer is image 1 of 2 total images of the two sheet source map set representing the eastern portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Pennsylvania South State Plane Coordinate System NAD27 (in Feet) (Fipszone 3702). All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. USGS maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 10 and 20 feet. Please pay close attention to map collar information on projections, spheroid, sources, dates, and keys to grid numbering and other numbers which appear inside the neatline. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.