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In eukaryotes, a first step towards the nuclear DNA compaction process is the formation of a nucleosome, which is comprised of negatively charged DNA wrapped around a positively charged histone protein octamer. Often, it is assumed that the complexation of the DNA into the nucleosome completely attenuates the DNA charge and hence the electrostatic field generated by the molecule. In contrast, theoretical and computational studies suggest that the nucleosome retains a strong, negative electrostatic field. Despite their fundamental implications for chromatin organization and function, these opposing views of nucleosome electrostatics have not been experimentally tested. Herein, we directly measure nucleosome electrostatics and find that while nucleosome formation reduces the complex charge by half, the nucleosome nevertheless maintains a strong negative electrostatic field. Our studies highlight the importance of considering the polyelectrolyte nature of the nucleosome and its impact on processes ranging from factor binding to DNA compaction.

The SNF2h remodeler slides nucleosomes most efficiently as a dimer, yet how the two protomers avoid a tug-of-war is unclear. Furthermore, SNF2h couples histone octamer deformation to nucleosome sliding, but the underlying structural basis remains unknown. Here we present cryo-EM structures of SNF2h-nucleosome complexes with ADP-BeFx that capture two potential reaction intermediates. In one structure, histone residues near the dyad and in the H2A-H2B acidic patch, distal to the active SNF2h protomer, appear disordered. The disordered acidic patch is expected to inhibit the second SNF2h protomer, while disorder near the dyad is expected to promote DNA translocation. The other structure doesn’t show octamer deformation, but surprisingly shows a 2 bp translocation. FRET studies indicate that ADP-BeFx predisposes SNF2h-nucleosome complexes for an elemental translocation step. We propose a model for allosteric control through the nucleosome, where one SNF2h protomer promotes asymmetric octamer deformation to inhibit the second protomer, while stimulating directional DNA translocation.

ISWI family chromatin remodeling motors use sophisticated autoinhibition mechanisms to control nucleosome sliding. Yet how the different autoinhibitory domains are regulated is not well understood. Here we show that an acidic patch formed by histones H2A and H2B of the nucleosome relieves the autoinhibition imposed by the AutoN and the NegC regions of the human ISWI remodeler SNF2h. Further, by single molecule FRET we show that the acidic patch helps control the distance travelled per translocation event. We propose a model in which the acidic patch activates SNF2h by providing a landing pad for the NegC and AutoN auto-inhibitory domains. Interestingly, the INO80 complex is also strongly dependent on the acidic patch for nucleosome sliding, indicating that this substrate feature can regulate remodeling enzymes with substantially different mechanisms. We therefore hypothesize that regulating access to the acidic patch of the nucleosome plays a key role in coordinating the activities of different remodelers in the cell. Every human cell contains nearly two meters of DNA, which is carefully packaged to form a dense structure known as chromatin. The building block of chromatin is the nucleosome, a unit composed of a short section of DNA tightly wound up around a spool-like core of proteins called histones. The tight structure of the nucleosome prevents the cell from accessing and ‘reading’ the genes in the packaged DNA, effectively switching off these genes. So the exact placement of nucleosomes helps manage which genes are turned on. Changing the position of the nucleosomes can ‘free’ the DNA and make genes available to the cell. Enzymes called chromatin remodelers move nucleosomes around – for example, they can make the histone core slide on the DNA strand. However, it is still unclear how these enzymes recognize nucleosomes. Previous research indicates that many proteins bind to nucleosomes by using a surface on the histone proteins called the acidic patch. Could chromatin remodelers also work by interacting with this acidic patch? To address this further, Gamarra et al. investigate how a chromatin remodeler enzyme known as SNF2h interacts with a nucleosome. By default, SNF2h is inactive because two of its regions called AutoN and NegC act as brakes. The experiments show that the acidic patch helps to bypass this inactivation and switches on SNF2h. Gamarra et al. propose that, when SNF2h docks on to the nucleosome, the patch provides a landing pad for the AutoN and NegC modules; this interaction activates the enzyme, which can then start remodeling the nucleosome. However, another type of chromatin remodeler also uses the patch to interact with nucleosomes but it does not have the AutoN and NegC regions. This suggests that chromatin remodelers work with the acidic patch in different ways. Overall, the findings deepen our understanding of how DNA is packaged in cells, and how this process may go wrong and cause disease.

The regulation of cell physiology depends largely upon interactions of functionally distinct proteins and cellular components. These interactions may be transient or long-lived, but often affect protein motion. Measurement of protein dynamics within a cellular environment, particularly while perturbing protein function with small molecules, may enable dissection of key interactions and facilitate drug discovery; however, current approaches are limited by throughput with respect to data acquisition and analysis. As a result, studies using super-resolution imaging are typically drawing conclusions from tens of cells and a few experimental conditions tested. We addressed these limitations by developing a high-throughput single-molecule tracking (htSMT) platform for pharmacologic dissection of protein dynamics in living cells at an unprecedented scale (capable of imaging >10 6 cells/day and screening >10 4 compounds). We applied htSMT to measure the cellular dynamics of fluorescently tagged estrogen receptor (ER) and screened a diverse library to identify small molecules that perturbed ER function in real time. With this one experimental modality, we determined the potency, pathway selectivity, target engagement, and mechanism of action for identified hits. Kinetic htSMT experiments were capable of distinguishing between on-target and on-pathway modulators of ER signaling. Integrated pathway analysis recapitulated the network of known ER interaction partners and suggested potentially novel, kinase-mediated regulatory mechanisms. The sensitivity of htSMT revealed a new correlation between ER dynamics and the ability of ER antagonists to suppress cancer cell growth. Therefore, measuring protein motion at scale is a powerful method to investigate dynamic interactions among proteins and may facilitate the identification and characterization of novel therapeutics.

Records of hydrologic parameters, especially those parameters that are directly linked to air temperature, were analyzed to find indicators of recent climate warming in Minnesota, USA. Minnesota is projected to be vulnerable to climate change because of its location in the northern temperate zone of the globe. Ice-out and ice-in dates on lakes, spring (snowmelt) runoff timing, spring discharge values in streams, and stream water temperatures recorded up to the year 2002 were selected for study. The analysis was conducted by inspection of 10-year moving averages, linear regression on complete and on partial records, and by ranking and sorting of events. Moving averages were used for illustrative purposes only. All statistics were computed on annual data. All parameters examined show trends, and sometimes quite variable trends, over different periods of the record. With the exception of spring stream flow rates the trends of all parameters examined point toward a warming climate in Minnesota over the last two or three decades. Although hidden among strong variability from year to year, ice-out dates on 73 lakes have been shifting to an earlier date at a rate of -0.13 days/year from 1965 to 2002, while ice-in dates on 34 lakes have been delayed by 0.75 days/year from 1979 to 2002. From 1990 to 2002 the rates of change increased to -0.25 days/year for ice-out and 1.44 days/year for ice-in. Trend analyses also show that spring runoff at 21 stream gaging sites examined occurs earlier. From 1964 to 2002 the first spring runoff (due to snowmelt) has occurred -0.30 days/year earlier and the first spring peak runoff -0.23 days/year earlier. The stream water temperature records from 15 sites in the Minneapolis/St Paul metropolitan area shows warming by 0.11 degrees C/year, on the average, from 1977 to 2002. Urban development may have had a strong influence. The analysis of spring stream flow rates was inconclusive, probably because runoff is linked as much to precipitation and land use as to air temperature. Ranking and sorting of annual data shows that a disproportionately large number of early lake ice-out dates has occurred after 1985, but also between 1940 and 1950; similarly late lake ice-in has occurred more frequently since about 1990. Ranking and sorting of first spring runoff dates also gave evidence of earlier occurrences, i.e. climate warming in late winter. A relationship of changes in hydrologic parameters with trends in air temperature records was demonstrated. Ice-out dates were shown to correlate most strongly with average March air temperatures shifting by -2.0 days for a 1 degrees C increase in March air temperature. Spring runoff dates also show a relationship with March air temperatures; spring runoff dates shift at a rate of -2.5 days/1 degrees C minimum March air temperature change. Water temperatures at seven river sites in the Minneapolis/St Paul metropolitan area show an average rise of 0.46 degrees C in river temperature/1 degrees C mean annual air temperature change, but this rate of change probably includes effects of urban development. In conclusion, records of five hydrologic parameters that are closely linked to air temperature show a trend that suggests recent climate warming in Minnesota, and especially from 1990 to 2002. The recent rates of change calculated from the records are very noteworthy, but must not be used to project future parameter values, since trends cannot continue indefinitely, and trend reversals can be seen in some of the long-term records. [PUBLICATION ABSTRACT]

Ghosts haunt Christina Rossetti's poetry. Amidst the lyrics, devotional poems, and children's verse, poems about ghosts and hauntings recur as material evidence of Rossetti's fascination with spectral presences. That fascination poses a particular interpretive puzzle in light of her religious convictions and piety. We might be tempted to identify the recurring ghosts as just another nineteenth-century flirtation with spiritualism – the spiritualism by which her brothers William and Gabriel were intrigued, attending séances and testing the validity of communications from the dead. Rossetti, however, clearly dismissed spiritualism as false belief and a means to sin. We might also be tempted to divide Rossetti's poetry into the secular and the sacred and to categorize the ghost poems as the former, yet much recent criticism on Rossetti has argued successfully for the pervasiveness of her religious voice even in works that seem not to be religious. Finally, in seeking to hear a religious resonance, we might be tempted to interpret her ghosts as representative of the Holy Ghost, yet that interpretation could only be asserted at the expense of the poems themselves; as narrative poems, most of them involve ghosts of dead lovers, desired by the living for themselves – not as experiences of God's presence. Rossetti's use of ghosts within short narrative or dialogic poems of the late 1850s and 60s concerning human desire for lost love invites closer inspection, especially when such poems overtly treat her religious beliefs.

:  A recent study by the Texas Bacteria Total Maximum Daily Load (TMDL) Task Force has recommended the use of load duration curves as a primary tool in calculating bacterial TMDLs. This method is attractive because it effectively integrates flow regimes into TMDL analyses, clearly communicates data through a method that is understandable to the general public, and has been successfully applied in TMDL studies in other states. To ease the creation of load duration curves, an automated load duration curve creation tool called LDCurve has been created within a Microsoft Excel framework. Web services and a webscraper are used to retrieve U.S. Geological Survey streamflow data and Texas Commission on Environmental Quality water quality data. Data are imported to the spreadsheet, combined to create flow and load duration curves, and plotted. Final steps result in a preliminary estimate of the overall load reductions needed to meet water quality standards in the modeled segment. LDCurve is currently only applicable in the state of Texas, but may be updated to model water quality throughout the nation using analogous web services from the EPA STORET database. By using automated data retrievals and computations, the LDCurve tool reduces the amount of time required to create curves and calculate load reductions to a matter of minutes. LDCurve and all supporting materials are available online for free download at: http://tools.crwr.utexas.edu/LDCurve/.

On this point, Deirdre David argues that Aurora's artistic power \"performs a 'service' for a patriarchal vision of the apocalypse\" and works against a materialist ideology that keeps human souls from God.16 Mermin, too, seems to acknowledge this quality when she describes the ending as combining \"in a transcendental unity\" Romney's socialism, Aurora's naturalism, and Barrett Browning's politics and spirituality, although she does not investigate the implicit contradiction within such a combination (p. 214). Herbert Tucker describes the formal pattern of Aurora Leigh as nested concentric rings, which place the text structurally within the epic tradition and within which Barrett Browning achieves great poetic freedom through the text's fluidity of tense and temporal position.18 As the concentric rings move from Italy to London and back again, and from Aurora's story to Marian Earle's to a treatise on poetry and back again, the text traces a path from division and separation to reconciliation.

In her apocalyptic poetry in particular, Rossetti employs this participatory hermeneutic to reject what Lynda Palazzo identifies as the Tractarians' \"renewed emphasis on woman's sinfulness, moral weakness and role in the Fall, which required the advent of a male saviour to redeem humanity.'\"' For the Victorian woman to attempt not only to interpret this oblique text but also to critique the gender ideology informing its archetypal female figures- the Woman Clothed with the Sun, the Whore of Babylon, the Bride of Christ- is to place herself at risk on multiple levels.

A recent study by the Texas Bacteria Total Maximum Daily Load (TMDL) Task Force has recommended the use of load duration curves as a primary tool in calculating bacterial TMDLs. This method is attractive because it effectively integrates flow regimes into TMDL analyses, clearly communicates data through a method that is understandable to the general public, and has been successfully applied in TMDL studies in other states. To ease the creation of load duration curves, an automated load duration curve creation tool called LDCurve has been created within a Microsoft Excel framework. Web services and a webscraper are used to retrieve U.S. Geological Survey streamflow data and Texas Commission on Environmental Quality water quality data. Data are imported to the spreadsheet, combined to create flow and load duration curves, and plotted. Final steps result in a preliminary estimate of the overall load reductions needed to meet water quality standards in the modeled segment. LDCurve is currently only applicable in the state of Texas, but may be updated to model water quality throughout the nation using analogous web services from the EPA STORET database. By using automated data retrievals and computations, the LDCurve tool reduces the amount of time required to create curves and calculate load reductions to a matter of minutes. LDCurve and all supporting materials are available online for free download at: http://tools.crwr.utexas.edu/LDCurve/.