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In order to become bioactive, proteins must be translated and protected from aggregation during biosynthesis. The ribosome and molecular chaperones play a key role in this process. Ribosome-bound nascent chains (RNCs) of intrinsically disordered proteins and RNCs bearing a signal/arrest sequence are known to interact with ribosomal proteins. However, in the case of RNCs bearing foldable protein sequences, no direct information is available on these interactions. Here, via a combination of chemical crosslinking and time-resolved fluorescence-anisotropy, we find that nascent chains of the foldable globin apoHmp1 − 140 interact with ribosomal protein L23 and have a freely-tumbling non-interacting N-terminal compact region comprising 63–94 residues. Longer RNCs (apoHmp1 − 189) also interact with an additional yet unidentified ribosomal protein, as well as with chaperones. Surprisingly, the apparent strength of RNC/r-protein interactions does not depend on nascent-chain sequence. Overall, foldable nascent chains establish and expand interactions with selected ribosomal proteins and chaperones, as they get longer. These data are significant because they reveal the interplay between independent conformational sampling and nascent-protein interactions with the ribosomal surface.

Many proteins must interact with molecular chaperones to achieve their native state in the cell. Yet, how chaperone binding-site characteristics affect the folding process is poorly understood. The ubiquitous Hsp70 chaperone system prevents client-protein aggregation by holding unfolded conformations and by unfolding misfolded states. Hsp70 binding sites of client proteins comprise a nonpolar core surrounded by positively charged residues. However, a detailed analysis of Hsp70 binding sites on a proteome-wide scale is still lacking. Further, it is not known whether proteins undergo some degree of folding while chaperone bound. Here, we begin to address the above questions by identifying Hsp70 binding sites in 2258 Escherichia coli (E. coli) proteins. We find that most proteins bear at least one Hsp70 binding site and that the number of Hsp70 binding sites is directly proportional to protein size. Aggregation propensity upon release from the ribosome correlates with number of Hsp70 binding sites only in the case of large proteins. Interestingly, Hsp70 binding sites are more solvent-exposed than other nonpolar sites, in protein native states. Our findings show that the majority of E. coli proteins are systematically enabled to interact with Hsp70 even if this interaction only takes place during a fraction of the protein lifetime. In addition, our data suggest that some conformational sampling may take place within Hsp70-bound states, due to the solvent exposure of some chaperone binding sites in native proteins. In all, we propose that Hsp70-chaperone-binding traits have evolved to favor Hsp70-assisted protein folding devoid of aggregation.

As the largest biogeochemically active terrestrial reserve of carbon (C), soils have the potential to either mitigate or amplify rates of climate change. Ecosystems with large C stocks and high rates of soil C sequestration, in particular, may have outsized impacts on regional and global C cycles. Montane meadows have large soil C stocks relative to surrounding ecosystems. However, anthropogenic disturbances in many meadows may have altered the balance of C inputs and outputs, potentially converting these soils from net C sinks to net sources of C to the atmosphere. Here, we quantified ecosystem-level C inputs and outputs to estimate the annual net soil C flux from 13 montane meadows spanning a range of conditions throughout the California Sierra Nevada. Our results suggest that meadow soils can be either large net C sinks (577.6 ± 250.5 g C m−2 y−1) or sources of C to the atmosphere (− 391.6 ± 154.2 g C m−2 y−1). Variation in the direction and magnitude of net soil C flux appears to be driven by belowground C inputs. Vegetation species and functional group composition were not associated with the direction of net C flux, but climate and watershed characteristics were. Our results demonstrate that, per unit area, montane meadows hold a greater potential for C sequestration than the surrounding forest. However, legacies of disturbance have converted some meadows to strong net C sources. Accurate quantification of ecosystem-level C fluxes is critical for the development of regional C budgets and achieving global emissions goals.

In order to become bioactive, proteins need to be biosynthesized and protected from aggregation during translation. The ribosome and molecular chaperones contribute to both of these tasks. While it is known that some ribosomal proteins (r-proteins) interact with ribosome-bound nascent chains (RNCs), specific interaction networks and their role within the ribosomal machinery remain poorly characterized and understood. Here, we find that RNCs of variable sequence and length (beyond the 1stC-terminal reside) do not modify the apparent stability of the peptidyl-transferase center (PTC) and r-proteins. Thus, RNC/r-protein interaction networks close to the PTC have no effect on the apparent stability of ribosome-RNC complexes. Further, fluorescence anisotropy decay, chemical-crosslinking and Western blots show that RNCs of the foldable protein apoHmp1-140have an N-terminal compact region (63–94 residues) and interact specifically with r-protein L23 but not with L24 or L29, at the ribosomal-tunnel exit. Longer RNCs bear a similar compact region and interact either with L23 alone or with L23 and another unidentified r-protein, or with molecular chaperones. The apparent strength of RNC/r-protein interactions does not depend on RNC sequence. Taken together, our findings show that RNCs encoding foldable protein sequences establish an expanding specific interaction network as they get longer, including L23, another r-protein and chaperones. Interestingly, the ribosome alone (i.e., in the absence of chaperones) provides indiscriminate support to RNCs bearing up to ca. 190 residues, regardless of nascent-chain sequence and foldability. In all, this study highlights the unbiased features of the ribosome as a powerful nascent-protein interactor.Significance StatementThe presence of interactions between nascent chains bearing a foldable amino-acid sequence (with no signal or arrest tags) and specific ribosomal proteins has never been experimentally demonstrated, up to now. Here, we identify the ribosomal protein L23 as a specific nascent-chain interacting partner. We show that L23 establishes noncovalent contacts with nascent chains of the multi-domain foldable model protein apoHmp, which lacks signal/arrest sequences. Interactions with another ribosomal protein and with the trigger-factor and Hsp70 chaperones were also detected. Interestingly, ribosomal-protein/nascent-chain complexes have similar apparent stability, in the case of nascent chains of variable sequence and degree of foldability. These findings are significant because they advance our knowledge on ribosome-mediated nascent-protein interaction networks and suggest avenues to prevent undesirable aggregation.

Correct protein folding is essential for the health and function of living organisms. Yet, it is not well understood how unfolded proteins reach their native state and avoid aggregation, especially within the cellular milieu. Some proteins, especially small, single-domain and apparent two-state folders, successfully attain their native state upon dilution from denaturant. Yet, many more proteins undergo misfolding and aggregation during this process, in a concentration-dependent fashion. Once formed, native and aggregated states are often kinetically trapped relative to each other. Hence, the early stages of protein life are absolutely critical for proper kinetic channeling to the folded state and for long-term solubility and function. This review summarizes current knowledge on protein folding/aggregation mechanisms in buffered solution and within the bacterial cell, highlighting early stages. Remarkably, teamwork between nascent chain, ribosome, trigger factor and Hsp70 molecular chaperones enables all proteins to overcome aggregation propensities and reach a long-lived bioactive state.

Though citizen science programs have been broadly successful in diverse scientific fields, their adoption has lagged in some disciplines, including soil science and ecosystem ecology. Collaborations with citizen scientists may be viewed as a conundrum in these disciplines, which often require substantial labor and technical experience; citizen scientists could improve sampling capacity but may reduce sample quality or require training and oversight prior to and while performing specialized tasks. To demonstrate the feasibility of incorporating citizen scientists into soil biogeochemistry research, we conducted a proof-of-concept study in high-elevation meadows of the Sierra Nevada in California. A collaboration between university researchers and citizen scientists allowed us to assess spatial and diel patterns of soil greenhouse gas (GHG) fluxes with an intensity and frequency that would otherwise be beyond the capacity of a typical research laboratory. This collaboration with citizen scientists increased our sampling intensity by over 700% while only doubling the sampling error relative to that of full-time researchers. With training and support from project scientists, citizen scientists collected data that demonstrate spatial independence of carbon dioxide, methane, and nitrous oxide at scales between 1 m and 175 m. Additionally, we found a lack of temporal variation over a 24-h period for all three GHGs. Citizen scientists participating in this one-day event reported levels of satisfaction commensurate with longer-term, immersive campaigns. The place-based event also proved an effective tool for teaching intangible concepts of soil biogeochemistry and promoting local conservation. Despite perceived barriers to entry, this study demonstrates the mutual benefits of citizen science collaborations in soil science and ecosystem ecology, encouraging adoption by disciplines that have been slow to take advantage of such collaborations. Short-term, local citizen science events can provide meaningful experiences for area residents and teach global biogeochemical cycles in a place-based context.

https://doi.org/10.15447/sfews.2017v15iss2art4Change in land use in agriculturally dominated areas is often assumed to provide positive benefits for land-owners and financial agricultural returns at the expense of biodiversity and other ecosystem services. For an agriculturally dominated area in the Central Valley of California we quantify the trade-offs among ecosystem services, biodiversity, and the financial returns from agricultural lands. We do this by evaluating three different landscape management scenarios projected to 2050 compared to the current baseline: habitat restoration, urbanization, and enhanced agriculture. The restoration scenario benefited carbon storage services and increased landscape suitability for birds, and also decreased ecosystem disservices (nitrous oxide emissions, nitrogen leaching), although there was a trade-off in slightly lower financial agricultural returns. Under the urbanization scenario, carbon storage, suitability for birds, and agricultural returns were negatively affected. A scenario which enhanced agriculture, tailored to the needs of a key species of conservation concern (Swainson’s Hawk, Buteo swainsoni), presented the most potential for trade-offs. This scenario benefitted carbon storage and increased landscape suitability for the Swainson's Hawk as well as 15 other focal bird species. However, this scenario increased ecosystem disservices. These spatially explicit results, generated at a scale relevant to land management decision-makers in the Central Valley, provide valuable insight into managing for multiple benefits in the landscape and an approach for assessing future land-management decisions.

Planning riparian restoration to resemble historic reference conditions requires an understanding of both local and regional patterns of plant species diversity. Thus, understanding species distributions at multiple spatial scales is essential to improve restoration planting success, to enhance long-term ecosystem functioning, and to match restoration planting designs with historic biogeographic distributions. To inform restoration planning, we examined the biogeographic patterns of riparian plant diversity at local and regional scales within a major western U.S.A. drainage, California's Sacramento-San Joaquin Valley. We analyzed patterns of species richness and complementarity (β-diversity) across two scales: the watershed scale and the floodplain scale. At the watershed scale, spatial patterns of native riparian richness were driven by herbaceous species, whereas woody species were largely cosmopolitan across the nearly 38,000 km 2 study area. At the floodplain scale, riparian floras reflected species richness and dissimilarity patterns related to hydrological and disturbance-driven successional sequences. These findings reinforce the importance of concurrently evaluating both local and regional processes that promote species diversity and distribution of native riparian flora. Furthermore, as restoration activities become more prevalent across the landscape, strategies for restoration outcomes should emulate the patterns of species diversity and biogeographic distributions found at regional scales. © 2011 Society for Ecological Restoration International.

Perennial pepperweed (Lepidium latifolium) is a potential threat to biodiversity and ecosystem function in the communities that it invades. The mechanism for its successful invasion of riparian and wetland environments includes reproduction via seed and root propagules and its ability to withstand long duration flooding and saline conditions once established. Controlling this species presents a number of challenges for land managers, including difficult property access, limited herbicide choice, and the varied success of weed control measures. In this study, we tested the efficacy of a nonchemical-modified tarp treatment, and compared posttreatment stem counts to herbicide treatments with Mow–glyphosate and chlorsulfuron in a wildland setting. We found that tarping applied in combination with a mow and till treatment was effective at controlling perennial pepperweed at levels similar to herbicide treatments with Mow–glyphosate and with Mow–chlorsulfuron. However, Mow–Till–Tarp treatment is extremely time consuming and has the potential to limit native plant community recovery.