Search

Introduction In 2013, the first successful European Citizens’ Initiative (ECI), called “Right2Water,” collected 1.9 million signatures. This broad European civil-society initiative demanded that the European Commission implement the human right to water and sanitation in European legislation. The campaign was organized by the European Federation of Public Service Unions (EPSU) and supported by a large number of NGOs and water activists throughout Europe. The Right2Water movement campaigned against the European Commission’s intention to further privatize drinking water utilities in Europe, following the UK example. The European campaign faced a large diversity of contexts in each Member State of the European Union. The state of affairs regarding public or private governance of water utilities and those utilities’ performance differed per country. Moreover, awareness about the theme of “the human right to water” among the general public, grassroots federations, NGOs, and trade unions was also quite diverse. Water in Europe is subject to both European and national law and responsibility is shared between the European Union and Member States, making it a suitable issue for an ECI. The initiative aimed to shift the European Commission’s focus from their market orientation to a rights-based, people-oriented water-policy approach (Right2Water, 2016). The Right2Water campaign joined in the ongoing struggle for water justice that, in divergent ways, was framed and organized by many civil-society groups, and took a stance against profit-driven water companies with the slogan “Water is a public good; not a commodity!" It urged the European Commission that water services in Europe should not be liberalized. From April 2012 to September 2013, the ECI “Right2Water” collected signatures across Europe. In 13 countries, the number of signatures passed the EU threshold, which was set by the European Commission for making such peoples’ initiatives mandatory to be discussed by the European Commission (see Table 12.1); the first successful ECI ever. The ECI is a tool established by the Lisbon Treaty (EC, 2009) as a means to bring the European Union closer to its citizens. It gives people an opportunity to bring an issue to the European political agenda if they manage to collect over one million signatures in one year’s time, from at least seven countries, with a minimum for each country (EC, 2011).

Water governance fundamentally deals with the question of how to organize decision-making about water access, use and management in contexts of diverging interests, conflicting normative repertoires, and unequal power relations. It aims to produce particular socio-natural orders by controlling water resources, infrastructure, investments, knowledge, truth, and ultimately, water users and authorities (Boelens, 2014; Bridge and Perreault, 2009). To achieve this, as the chapters in Part I have illustrated, water governance reforms and interventions commonly emphasize strongly de-politicized common wellbeing, shared progress, and efficient, rational resource management. This naturalizing discourse of sustainable, progressive, clean development, achieved by egalitarian “stakeholders,” obscures the fact that water reforms and interventions entail competing claims and conflicts over water, territorial ordering, and reconfiguring socio-economic and politico-cultural realities (Harris and Alatout, 2010; Hommes et al., 2016; Kaika, 2006). These issues are directly related to disputes over problem definitions, knowledge frameworks, ontological meanings, decision-making powers and preferred solutions. The chapters in Part II will focus on this field of water (in)justice, the dynamics of contested imaginaries and materializing socio-natural, techno-political networks: de-patterning and re-patterning multi-scalar hydrosocial territories (Baviskar, 2007; Boelens et al., 2016; Linton and Budds, 2014; Swyngedouw, 2004, 2009). A governmentality perspective toward disputed, overlapping, hybridizing hydrosocial territories may help us to understand how water control is embedded in the broader political context of governance over and through socio-natures. Governmentality refers to the ways societies are governed, not only through direct application of laws and military force, but also through subtle and invisible “capillary” working of power to control, at once, the conduct of people and their socio-environment (Foucault, 1991). The concept of hydrosocial territory provides a lens to analyze water flows, water infrastructures, and water control as simultaneously, interactively constituted compositions of physical, social, political and symbolic entities and dimensions. Together, these domains form multi-scaled, networked mosaics, which produce techno-political and socio-ecological territoriality (Boelens et al., 2016; cf. Hommes et al., 2016; Hoogesteger et al., 2016; Swyngedouw, 2007). This analytical framework views humans and nature - social, technical and natural - not as separate entities that interact, but as mutually influencing, co-producing and constituting each other, in complex ways (see also Harris and Alatout, 2010; Latour, 1994).

Introduction For the last couple of decades, groundwater tables in the Saïss agricultural plain, mid-north Morocco, have been dropping. This is partly due to intensified groundwater use in agriculture and changing cropping patterns. Specifically, since the 1980s, farmers have increasingly dug wells to engage in commercial agriculture. From growing rain-fed crops, they have shifted to higher-value irrigated crops, notably onions and other vegetables. Changing tenure relations (land privatization, most notably) and new agricultural policies that actively promoted modernization have further intensified groundwater use. Today, water-intensive crops produced in the region are sold on regional, national and international markets. They produce new material and cultural linkages between hitherto-unconnected life worlds, interactively reshaping socio-natural relations. They open up new possibilities of living and being, but also close off old ones. In this chapter, we describe and analyze these changes to interrogate them from a feminist-justice perspective. We use material from extensive ethnographic fieldwork, conducted over a two-year period from 2011 to 2013, complemented by various recurring several-week visits over 2014 and 2015. We have focused on anecdotes regarding the changing groundwater and irrigation dynamics that illustrate how they are intimately linked to changing gender relations. Rather than merely assessing or mapping the gendered impacts of intensifying groundwater use, our purpose is to tease out how prevailing gendered ways of being, working, and relating co-shape and in turn are shaped by new agricultural and groundwater dynamics, in ways that are not always straightforward or easy to predict. Hence, our argument is that groundwater abstraction and gender relations dynamics co-constitute each other, which also means that one cannot be comprehended without the other. We show this by focusing on how intensifying groundwater use, and the larger changes it provokes, is made possible through and shaped by changing the gendered organization of labor and space. Contemporary agrarian policies in Morocco intended to promote a new kind of green revolution are significantly premised on promoting a particular gendered organization of farming (and consequently of families), souting specific roles, responsibilities and rightsto men and to women. Hence, the two main protagonists of agricultural modernization that the Moroccan government focuses on and wants to support are new agricultural investors, and those existing farmers who have the spirit and willingness to modernize. In public policies, these protagonists are clearly cast as men - with farms figuring as enterprises that operate with a logic of profit maximization.

Standing Rock The protesters called themselves “water protectors.” They came from all corners of the US and Canada, and included members of over 100 Native American tribes and their non-indigenous allies. At the peak of the protest, in early autumn 2016, they numbered in the thousands, but by late February, when the Governor sent some 200 police officers to clear the encampment, only a few dozen remained. Those who left burned their temporary shelters in an act of defiance, and the embers were still smoldering when the police moved in and arrested the few who stayed behind (Smith and Blinder, 2017). For a brief period, it seemed the protesters might have won: the Obama administration halted construction of the Dakota Access pipeline and asked the US Army Corps of Engineers, which was overseeing the project, to find an alternate route. But then the political winds shifted. A right-wing populist was elected President and the winter cold brought with it a form of revanchist politics that had little patience for Native Americans and their atavistic attachments to land and water (Healy and Fandos, 2016; Smith, 2017). The protesters had gathered on the banks of the Cannonball River, near its confluence with the Missouri, where the Texas-based Energy Transfer Partners planned to build the last section of the pipeline. Construction was nearly complete, and all that remained was a short stretch to be built under the Missouri River. The pipeline route consisted entirely of private land, but because it had to cross a navigable waterway, it required the approval of the US Army Corps of Engineers (New York Times, 2016a). Despite opposition from the US Environmental Protection Agency, the US State Department and the Sioux Nation’s Tribal Historic Preservation Office, the Army Corps seemed determined to fast-track the pipeline’s approval. Contrary to federal requirements, it approved construction without meaningful consultation with the Standing Rock Sioux, whose lands and waters the pipeline would impact (Archambault, 2016; Sammon, 2016). Although the pipeline would not cross the Standing Rock reservation, instead passing half a mile north of the reservation boundary, tribal members argued that it crossed land they considered sacred (and which had formerly been part of Sioux territory, of which they were dispossessed in the 1870s when they were confined to the reservation).

The nucleotide excision repair protein complex ERCC1-XPF is required for incision of DNA upstream of DNA damage. Functional studies have provided insights into the binding of ERCC1-XPF to various DNA substrates. However, because no structure for the ERCC1-XPF-DNA complex has been determined, the mechanism of substrate recognition remains elusive. Here we biochemically characterize the substrate preferences of the helix-hairpin-helix (HhH) domains of XPF and ERCC-XPF and show that the binding to single-stranded DNA (ssDNA)/dsDNA junctions is dependent on joint binding to the DNA binding domain of ERCC1 and XPF. We reveal that the homodimeric XPF is able to bind various ssDNA sequences but with a clear preference for guanine-containing substrates. NMR titration experiments and in vitro DNA binding assays also show that, within the heterodimeric ERCC1-XPF complex, XPF specifically recognizes ssDNA. On the other hand, the HhH domain of ERCC1 preferentially binds dsDNA through the hairpin region. The two separate non-overlapping DNA binding domains in the ERCC1-XPF heterodimer jointly bind to an ssDNA/dsDNA substrate and, thereby, at least partially dictate the incision position during damage removal. Based on structural models, NMR titrations, DNA-binding studies, site-directed mutagenesis, charge distribution, and sequence conservation, we propose that the HhH domain of ERCC1 binds to dsDNA upstream of the damage, and XPF binds to the non-damaged strand within a repair bubble.

GH (growth hormone) binding to the GHR (GH receptor) triggers essential signalling pathways that promote growth and metabolic regulation. The sensitivity of the cells to GH is mainly controlled by the endocytosis of the receptor via βTrCP (β-transducin repeat-containing protein). In the present study, we show that βTrCP interacts directly via its WD40 domain with the UbE (ubiquitin-dependent endocytosis) motif in GHR, promoting GHR ubiquitination in vitro. NMR experiments demonstrated that the UbE motif is essentially unstructured, and, together with functional mapping of the UbE and βTrCP WD40 residues necessary for binding, led to a unique interaction model of βTrCP with GHR–UbE. This interaction is different from the conventional βTrCP–substrate interactions described to date. This interaction therefore represents a promising specific target to develop drugs that inhibit GHR endocytosis and increase GH sensitivity in cachexia patients.

Conotoxins are small peptides present in the venom of cone snails. The snail uses this venom to paralyze and capture prey. The constituent conopeptides display a high level of chemical diversity and are of particular interest for scientists as tools employed in neurological studies and for drug development, because they target with exquisite specificity membrane receptors, transporters, and various ion channels in the nervous system. However, these peptides are known to contain a high frequency and variability of post-translational modifications—including sometimes O-glycosylation—which are of importance for biological activity. The potential application of specific conotoxins as neuropharmalogical agents and chemical probes requires a full characterization of the relevant peptides, including the structure of the carbohydrate part. In this review, the currently existing knowledge of O-glycosylation of conotoxins is described.

For many Andean communities, irrigation systems form the basis for accessing water for agricultural production and, as such, they are an important axis around which collective action is mobilized (Bolin, 1990; Hoogesteger, 2013b; Verzijl and Guerrero Quispe, 2013). In the Andean context, with unpredictable climates, unstable geophysical conditions, and changing irrigation policies, more than almost any other economic activity, irrigation is grounded in collective action that is based on mutual dependence and intensive cooperation among users. In the Andes, besides privately managed irrigation systems owned by landlords and agribusiness companies, two forms of irrigation development and water use systems prevail, which in broad terms can be divided into state led and community based. State-led irrigation development has been characterized by its large scale, high cost, market-oriented and top-down management approach since at least the 1960s. On the other hand, community-managed irrigation systems tend to be small scale, constructed with local resources and know-how, managed through collective action from the bottom up and often with a very diverse production rationale. These two forms of irrigation management have grown closer to each other in the last three decades as on the one side collective action has come to play a more important role in formerly state-managed irrigation systems and, on the other, the state has become more active in funding the modernization of community-managed irrigation systems.

In 1996, Minera San Xavier (MSX), a Mexican tributary of the Canadian mining company Newgold Inc., announced it wanted to start a large open-pit gold and silver mine in the municipality of Cerro de San Pedro, in the state of San Luis Potosí. The mining site occupies 373 hectares of ejido 2 community land. This was highly controversial as the scale and type of the mining operation would impose a heavy burden on the available land and water and generate many adverse social and environmental impacts. Resistance was fierce, and several opposition groups united to form the Frente Amplio Opositor (Broad Opposition Front; BOF). Despite the opposition, however, MSX started operations in 2007. To date, its presence is still disputed.

Water is and has always been the “life stream” of rural livelihoods in Andean communities. Access to water has for centuries been guaranteed through various forms of collective action and autonomous governance structures (Boelens, 2015). Until now, collective action has assured individual water access and is mostly based on local, inter- and intra-community water rights systems that shape, and are shaped by, water flows and infrastructure, local water-related practices, authorities and territory, and particular world views on how societies relate to water and nature (see Beccar et al., 2002; Hoogesteger et al., 2016).

Structure determination of protein-nucleic acid complexes remains a challenging task. Here we present a simple method for generating crystals of a CsrA-nucleic acid complex, guided entirely by results from nuclear magnetic resonances spectroscopy (NMR) spectroscopy. Using a construct that lacks thirteen non-essential C-terminal residues, efficient binding to DNA could be demonstrated. One CsrA dimer interacts with two DNA oligonucleotides, similar to previous findings with RNA. Furthermore, the NMR study of the CsrA-DNA complex was the basis for successfully homing in on conditions that were suitable for obtaining crystals of the CsrA-DNA complex. Our results may be useful for those cases where RNA in protein-nucleic acid complexes may be replaced by DNA.

This review of pollution in the open oceans updates a report on this topic prepared by GESAMP five years previously (Reports and Studies No. 79, GESAMP, 2009). The latter report, the first from GESAMP focusing specifically on the oceans beyond the 200 m depth contour, was prepared for purposes of the Assessment of Assessments, the preparatory phase of a regular process for assessing the state of the marine environment, led jointly by the United Nations Environment Programme (UNEP) and the Intergovernmental Oceanographic Commission (UNESCO-IOC).

International audience; The glycopeptide CcTx, isolated from the venom of the piscivorous cone snail Conus consors, belongs to the κA-family of conopeptides. These toxins elicit excitotoxic responses in the prey by acting on voltage-gated sodium channels. The structure of CcTx, a first in the κA-family, has been determined by high-resolution NMR spectroscopy together with the analysis of its O-glycan at Ser7. A new type of glycopeptide O-glycan core structure, here registered as core type 9, containing two terminal L-galactose units {α-L-Galp-(1→4)-α-D-GlcpNAc-(1→6)-[α-L-Galp-(1→2)-β-D-Galp-(1→3)-]α-D-GalpNAc-(1→O)}, is highlighted. A sequence comparison to other putative members of the κA-family suggests that O-linked glycosylation might be more common than previously thought. This observation alone underlines the requirement for more careful and in-depth investigations into this type of post-translational modification in conotoxins.

The tricarboxylic acid citric acid is a key intermediary metabolite in organisms from all domains of the tree of life. Surprisingly, this metabolite specifically interacts with the light-induced signaling state of the photoactive yellow protein (PYP), such that, at 30 mM, it retards recovery of this state to the stable ground state of the protein with up to 30%, in the range from pH 4.5 to pH 7. We have performed a detailed UV/vis spectroscopic study of the recovery of the signaling state of wild type (WT) PYP and two mutants, H108F and Delta25-PYP, derived from this protein, as a function of pH and the concentration of citric acid. This revealed that it is the dianionic form of citric acid that binds to the pB state of PYP. Its binding site is located in between the N-terminal cap and central beta-sheet of PYP, which is accessible only in the signaling state of the protein. The obtained results show how changes in the distribution of subspecies of the signaling state of PYP influence the rate of ground state recovery.

We summarize the use of NMR spectroscopy in the production and the screening of stability and foldedness of protein domains, and apply it to the RING domains of E3 ubiquitin-ligases. RING domains are involved in specific interactions with E2 ubiquitin-conjugating enzymes and thus play an essential role in the ubiquitination pathway. Protein production of the Zn(2+) containing and cysteine rich RING domains for molecular studies frequently turns out to be problematic. We compared the expression and solubility of 14 E3 RING/U-box domains fused to the N-terminal tags of His(6), His(6)-GB1, His(6)-Trx and His(6)-GST at small scale and analyzed, by NMR spectroscopy, their correct folding after purification. The addition of GST, Trx or GB1 to the N-terminal His(6) tag significantly improved both the expression and solubility of target proteins as compared to His(6) tag alone. More importantly most of the immobilized metal affinity chromatography (IMAC) purified proteins were largely unfolded as judged by analysis of the (1)H-(15)N HSQC spectra. We demonstrate that imidazole causes a concentration dependent decrease in stability of RING proteins ascribed to metal depletion and resulting in unfolding or precipitation. In contrast, using glutathione affinity chromatography, the His(6)-GST fused RING and U-box domains were purified as correctly folded proteins with high yields. Our data clearly demonstrate that IMAC should be avoided and that GST-fusion affinity chromatography is generally applicable for expression and purification of Zn(2+) containing proteins.

Recent technological advances in NMR spectroscopy have alleviated the size limitations for the determination of biomolecular structures in solution. At the same time, novel NMR parameters such as residual dipolar couplings are providing greater accuracy. As this review shows, the structures of protein-protein and protein-nucleic acid complexes up to 50 kDa can now be accurately determined. Although de novo structure determination still requires considerable effort, information on interaction surfaces from chemical shift perturbations is much easier to obtain. Advances in modelling and data-driven docking procedures allow this information to be used for determining approximate structures of biomolecular complexes. As a result, a wealth of information has become available on the way in which proteins interact with other biomolecules. Of particular interest is the fact that these NMR-based methods can be applied to weak and transient protein-protein complexes that are difficult to study by other structural methods.

Three-dimensional domain swapping is a common phenomenon in pancreatic-like ribonucleases. In the aggregated state, these proteins acquire new biological functions, including selective cytotoxicity against tumour cells. RNase A is able to dislocate both N- and C-termini, but usually this process requires denaturing conditions. In contrast, bovine seminal ribonuclease (BS-RNase), which is a homo-dimeric protein sharing 80% of sequence identity with RNase A, occurs natively as a mixture of swapped and unswapped isoforms. The presence of two disulfides bridging the subunits, indeed, ensures a dimeric structure also to the unswapped molecule. In vitro, the two BS-RNase isoforms interconvert under physiological conditions. Since the tendency to swap is often related to the instability of the monomeric proteins, in these paper we have analysed in detail the stability in solution of the monomeric derivative of BS-RNase (mBS) by a combination of NMR studies and Molecular Dynamics Simulations. The refinement of NMR structure and relaxation data indicate a close similarity with RNase A, without any evidence of aggregation or partial opening. The high compactness of mBS structure is confirmed also by H/D exchange, urea denaturation, and TEMPOL mapping of the protein surface. The present extensive structural and dynamic investigation of (monomeric) mBS did not show any experimental evidence that could explain the known differences in swapping between BS-RNase and RNase A. Hence, we conclude that the swapping in BS-RNase must be influenced by the distinct features of the dimers, suggesting a prominent role for the interchain disulfide bridges.

Item does not contain fulltext

1H NMR relaxation is used to study the self-assembly of a double thermoresponsive diblock copolymer in dilute aqueous solution. Above the first transition temperature, at which aggregation into micellar structures is observed, the trimethylsilyl (TMS)-labeled end group attached to the shell-forming block shows a biphasic T-2 relaxation. The slow contribution reflects the TMS groups located at the periphery of the hydrophilic shell, in agreement with a star-like micelle. The fast T-2 contribution corresponds to the TMS groups, which fold back toward the hydrophobic core, reflecting a flower-like micelle. These results confirm the formation of block copolymer micelles of an intermediate nature (i.e., of partial flower-like and star-like character), in which a part of the TMS end groups folds back to the core due to hydrophobic interactions. image