Every Cubs fan is ten years old tonight.

For the first time since 1945, the Cubs are in a World Series.  They haven't won one since 1908.  In that year, Harriet Tubman was still alive.  So was Mark Twain.  And Leo Tolstoy. And Geronimo. And Butch Cassidy and the Sundance Kid.

My high school was founded that year.

It's just baseball, yes, and I have philosophical objections to the culture of professional sports in this country and elsewhere.

But let me repeat: in this, the Year of Our Lord Two Thousand and Sixteen, the Cubs are in the World Series. For all the years since '08, for all the years they had great players like Ernie Banks and Billy Williams and Ron Santo and Ryne Sandberg, this time, they finally, finally made it. 

For fans who remember 1945 and bovid mammals of the genus Capra; for those who recall the implosions of 1969 and 1984 and 2003, this time, they did it (Yeah, there were a couple other times they were in the playoffs since '84, but those never felt like their year).  This time, they finally did it.

I remember, in the 70s, my mother taking me and my brother and our friends to the Cubs games, especially on Fridays, which was Ladies' Day and tickets were cheap (Fridays didn't become popular until the 80s).  It was the era of Reggie Jackson and the Pirates and the Big Red Machine of Cincinnati.

There were peanuts outside and hot dogs inside and vendors selling Old Style beer (which of course we didn't drink). There were Andy Frain ushers and no one had even thought about putting seats on the roofs of buildings across the street. There was that deliciously analog giant scoreboard in center field.  (And, of course, there were no big screen high definition TVs.).

I remember our neighbor's cousin from Japan coming to see a baseball game in America and being wowed by Wrigley Field.

I remember some of the coldest spring days of my life sitting along the unreserved seats of left field.

I remember when the Cubs scored 22 runs in a single game. And lost. (I also remember when WGN replayed the game just because it was so awesome.).

I remember when they installed lights and being relieved they architecturally matched the stadium.

I remember commemorating the 50th anniversary of my high school's new building and the seventy-fifth anniversary of its founding by walking down Addison to see the Cubs play.

And I remember when Hank Aaron came for his first appearance in Chicago after breaking Babe Ruth's home run record and the entire stadium stood and gave him a standing ovation even though he was on the other team. 

I remember Jack Brickhouse and being kind of disappointed when he retired and was replaced by Harry Caray who, of course, had covered the White Sox, which was just wrong.

I remember friends being fans of the Reds and the Pirates and the Dodgers and the White Sox, which was just wrong. (For the record, I was not opposed to their winning the World Series a couple years back.  I don't like their new stadium, though.  Or the fact that they took Comiskey off the name).

I totally shouldn't care about professional athletes making millions for their billionaire employers for mediocre performances over the course of a century.  And part of me doesn't.

But it's the Cubs.  And today, every Chicagoan who remembers is ten years old again.  And tonight, that's sublime.

Even if they don't beat Cleveland.  But they will. Unless they don't.  In which case they will do so in the most heart-breakingly way possible. Because they're the Cubs.

And it's what they do.

And there will be a next year.

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Note, this offer is for Current/Existing Game Pass Ultimate Members only. Expired/New users can redeem up to 3-Years worth of Xbox Live Gold codes and convert their Xbox Live Gold membership to 2-Years of Game Pass Ultimate by joining Ultimate Membership. (3:2 Ratio)
 
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Melon Game via Eneba has 13-Months Xbox Game Pass Ultimate Subscription via Xbox Live Gold Conversion (Digital Delivery) on sale starting from ~$56 when you follow the instructions below.

• Note: Offer valid for New or Expired Xbox Game Pass Ultimate subscribers only. For current/active Game Pass Ultimate Members, check this thread.

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Deal Instructions:

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Half off select accessories via verizon up. Elite 2 controller 89.99

 

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Eneba offers 50-Day of Xbox Game Pass Ultimate via Xbox Game Pass Core Conversion for around $11.05 when you follow the instructions below.

• Note, this offer is for Current/Existing Game Pass Ultimate Members only.

Deal Instructions:

• Click here to visit the product page.
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• Select payment method PayPal/Credit Card (service fees will be applied here)
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• Click here to redeem your 3-Month Xbox Live Gold Membership code(s).
• You will be prompted to extend your Game Pass Ultimate by 50 days. The renewal date should be the same as your Xbox Game Pass Ultimate plus 50 days (Maximum 36 months).

About Game Pass Ultimate:

• Xbox Game Pass Ultimate Membership includes Xbox Live Gold and Xbox Game Pass with over 100 high-quality games to play with friends on console, PC, phones, and tablets, plus an EA Play membership, all for one low price.

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Ibiza, a stunning island in the Mediterranean Sea, is globally renowned for its electric nightlife and vibrant party scene. This picturesque island has become the epitome of summer party destinations. It attracts visitors from all corners of the world with its beautiful beaches, mesmerizing sunsets, and, most notably, its unparalleled nightlife. Clubs in Ibiza are not just venues; they are the beating heart of the island’s culture as they showcase world-class DJs, groundbreaking music, and unforgettable experiences to define the essence of a perfect summer getaway. The purpose of this article is to guide you through the exhilarating nightlife of Ibiza. Whether you’re a seasoned club-goer or a first-time visitor, this comprehensive guide to the top 10 must-visit nightclubs will ensure you hit the hottest spots and make the most of your nights in Ibiza. From iconic institutions to cutting-edge clubs, we’ve curated a list that captures the diversity and vibrancy of Ibiza’s nightlife. These places promise unforgettable nights filled with music, dance, and celebration. Why Ibiza? Ibiza stands as a beacon for nightlife enthusiasts from around the globe. It’s celebrated for its unmatched party scene and vibrant atmosphere. But what exactly catapults this Balearic Island into the spotlight as a world-renowned destination for nightlife? Diverse Music Scene: Ibiza’s music landscape is as varied as it is electrifying. From techno and house to EDM and indie rock, the island offers something for every taste, hosted across an array of clubs and venues. Vibrant Atmosphere: There’s an infectious energy that permeates Ibiza’s nightlife. Whether you’re dancing under the stars at an outdoor beach club or losing yourself in the immersive environment of an underground venue, the atmosphere is always charged with excitement. Stunning Venues: Ibiza’s nightclubs are more than just places to dance; they’re architectural wonders and sensory experiences. From clubs with breathtaking views of the Mediterranean to venues that push the boundaries of interior design, Ibiza offers stunning settings that elevate the party experience to new heights. Criteria for Selection When it comes to pinpointing the top 10 nightclubs in Ibiza, our selection is driven by a carefully considered set of criteria. We aim to showcase venues that not only dominate the nightlife scene but also offer unique experiences that leave a lasting impression. The following factors play a crucial role in our selection: Music Genres: A club’s musical offerings are pivotal. We look for diversity and quality, favoring venues that host world-renowned DJs to suit a wide range of musical tastes. Atmosphere: The vibe of a nightclub is its soul. We seek out places that offer more than just a dance floor. In fact, we prioritize those who create an immersive experience through their energy, design, and crowd.  Reputation: A club’s reputation is built on years of delivering exceptional nights out. We consider the esteem in which venues are held by both locals and international visitors, as well as their recognition within the global nightlife community. Overall Experience: Finally, we factor in the overall experience, including the quality of sound and lighting systems, the presence of unique features (like outdoor terraces or poolside dance floors), and the level of service provided. A club that offers an unforgettable night out, from entry to exit, makes our list.  Club 1: Amnesia Ibiza Location San Rafael, Ibiza Music Genres Techno House EDM Atmosphere Amnesia Ibiza stands out for its high-energy ambiance which makes it a sanctuary for dance music enthusiasts. The club’s iconic terrace offers an immersive experience that draws partygoers into a world of euphoric beats and rhythms. Notable Features Famous Foam Parties: These unique and wildly entertaining parties are a signature Amnesia experience. State-of-the-Art Sound System: Delivers crystal-clear audio that enhances the musical journey. Spectacular Light Shows: Visually stunning displays complement the sonic experience, creating unforgettable moments.  Highlights Legends of the electronic music scene, such as Carl Cox and Marco Carola, are regulars here, guaranteeing an unmatched lineup of performances. Club 2: Pacha Ibiza Location Ibiza Town, Ibiza Music Genres House Techno EDM Atmosphere Pacha Ibiza exudes a glamorous and chic vibe and offers a luxurious nightclub experience. Its atmosphere attracts a stylish crowd to dance the night away in one of the island’s most iconic settings. Notable Features Iconic Cherry Logo: A symbol of the club’s heritage and prestige. VIP Sections: For those seeking a more exclusive experience, with stunning views of Ibiza Town. Elegant Decor: Pacha’s interior design combines style with comfort, creating a uniquely sophisticated environment. Highlights Hosting world-renowned DJs like David Guetta and Martin Solveig, Pacha remains at the forefront of Ibiza’s nightlife scene. Club 3: DC-10 Location Sant Josep de sa Talaia, Ibiza Music Genres Techno House Atmosphere DC-10 is celebrated for its raw and underground atmosphere. This club offers an intimate setting where the focus is squarely on the music and the communal experience of dance. Notable Features Open-Air Terrace: Provides a unique outdoor clubbing experience. Industrial Vibe: The club’s minimalist, warehouse-style setup enhances its gritty, authentic feel. Immersive Experiences: DC-10’s events are designed to create a deep connection between the DJs and the crowd. Highlights Home to the legendary Circoloco parties, DC-10 attracts top DJs such as Seth Troxler and The Martinez Brothers to ensure an unparalleled lineup and vibe. Club 4: Ushuaïa Ibiza Beach Club Location Playa d’en Bossa, Ibiza Music Genres EDM House Atmosphere Ushuaïa Ibiza Beach Club is the epitome of an extravagant outdoor party experience. With its poolside setting, it invites guests to dance under the sun and stars in one of the most breathtaking venues on the island. Notable Features Massive Stage: Hosts some of the biggest names in the music industry. Stunning Visuals: Offers a sensory feast with its impressive light shows and stage productions. Celebrity Appearances: A hotspot for celebrities, adding to the club’s allure. Highlights Iconic pool parties featuring world-class performances by artists like Calvin Harris and Avicii are the heart of Ushuaïa’s appeal. Club 5: Hï Ibiza Location Playa d’en Bossa, Ibiza Music Genres Techno House EDM Atmosphere Hï Ibiza offers a sophisticated and futuristic nightclub experience. Its immersive atmosphere is crafted through cutting-edge sound and stunning visuals that make every night a journey into the future of nightlife. Notable Features Cutting-Edge Sound System: Delivers unparalleled audio quality. Immersive Production: Engages all senses for a complete clubbing experience. Stunning Visuals: The club’s design and lighting create a mesmerizing environment. Highlights Glitterbox parties at Hï Ibiza shine with disco and house music vibes, featuring top DJs and a vibrant, inclusive atmosphere. Club 6: Privilege Ibiza Location San Rafael, Ibiza Music Genres House Techno EDM Atmosphere Privilege Ibiza is recognized for its massive scale and extravagant atmosphere. Holding the Guinness World Record for the world’s largest club, it offers an immersive experience with panoramic views of Ibiza. Notable Features World’s Largest Club: A title that adds to its legendary status. Panoramic Views: The club’s location and design provide breathtaking views of the island. Highlights Resistance parties at Privilege are a highlight, featuring top techno and house DJs like Adam Beyer and Charlotte de Witte to create unforgettable nights. Club 7: Eden Ibiza Location San Antonio, Ibiza Music Genres House Techno EDM Atmosphere Eden Ibiza strikes a perfect balance between lively and intimate as it creates an energetic atmosphere for music lovers to enjoy themselves up close with the performers. Notable Features Renovated Venue: A recent makeover has injected new life into Eden, enhancing its appeal. Impressive LED Screens: Add to the visual experience, complementing the beats with captivating displays. Intimate Dance Floor: Despite its size, Eden maintains an intimate feel, making every night more personal. Highlights Toolroom parties at Eden are a standout, with house music legends like Mark Knight behind the decks. Club 8: Heart Ibiza Location Ibiza Town, Ibiza Music Genres Eclectic Avant-garde Live performances Atmosphere Heart Ibiza offers an artistic and immersive experience with a blend of music, art, and gastronomy into a unique nightlife adventure to tantalize all senses. Notable Features Fusion of Music, Art, and Gastronomy: A unique concept that stands out in Ibiza’s nightlife. Stunning Interior Design: Adds to the immersive experience with its beauty and creativity. Highlights Heart Factory events are particularly notable for their live performances and DJ sets by international artists, making each night a unique journey. Club 9: Ibiza Rocks Hotel Location San Antonio, Ibiza Music Genres Indie Rock EDM Atmosphere Ibiza Rocks Hotel is known for its lively and vibrant environment, where the poolside setting adds a refreshing twist to the traditional nightclub experience. Notable Features Outdoor Stage: Offers a unique venue for live music under the Ibiza sun and stars. Pool Parties: Bring an energetic and fun atmosphere, perfect for daytime reveling. Live Music Events: Unlike many other clubs focusing solely on DJs, Ibiza Rocks hosts live bands and performances. Highlights The Ibiza Rocks Pool Party series is a summer highlight, featuring an eclectic mix of live performances by indie bands and DJs. It offers something different from the typical club scene. Club 10: Octan Ibiza Location Playa d’en Bossa, Ibiza Music Genres Techno House Atmosphere Octan Ibiza brings a raw and underground vibe to the forefront of Ibiza’s nightlife. Its intimate setting allows for an immersive clubbing experience that’s both intense and personal. Notable Features Intimate Dance Floors: Designed to create a close connection between the DJs and the crowd. Immersive Experiences: Focused on delivering cutting-edge underground parties. Legacy: Carrying the torch from the famed Sankeys Ibiza, Octan continues to push the boundaries of the electronic music scene. Highlights The club is a haven for enthusiasts of techno and house music, and it hosts cutting-edge underground parties with leading DJs in the genre. Frequently Asked Questions (FAQ) What is the legal drinking age in Ibiza? The legal drinking age in Ibiza is 18 years old. Is there a dress code for Ibiza nightclubs? Dress codes can vary. While some clubs are more relaxed, others might require a more upscale attire. It’s best to check the specific club’s policy before going. How much does it cost to enter Ibiza nightclubs? Entrance fees can range widely from club to club and depending on the event or DJ playing. Prices can vary from €20 to over €50. VIP services and tables will cost significantly more. Are there any age restrictions for entry into nightclubs? Yes, most nightclubs in Ibiza have an age restriction of 18 years and above. Is it necessary to book tickets in advance for Ibiza nightclubs? It’s highly recommended to book tickets in advance, especially for popular events, to ensure entry and often at a lower price. What are the typical opening hours for Ibiza nightclubs? Nightclubs usually open around midnight and can go on till dawn, with peak hours from 1 AM to 4 AM. Are there any safety tips for clubbing in Ibiza? Stay hydrated, especially if dancing for long periods. Keep an eye on your belongings. Never accept drinks from strangers. Stick with your friends and plan how to get back to your accommodation safely. Conclusion We’ve explored the top 10 must-visit nightclubs in Ibiza, and each offers its unique vibe. From the extravagant and glamorous to the raw and underground, Ibiza’s nightlife is unparalleled. Its diverse music scenes and world-renowned DJs provide an unforgettable experience for every partygoer. Whether you’re drawn to the legendary foam parties of Amnesia, the sophisticated atmosphere of Hï Ibiza, or the intimate underground vibes of Octan, the island caters to all. We encourage you to dive into the vibrant energy of Ibiza’s nightlife scene and experience the magic of these iconic clubs firsthand.  

The post Top 10 Must-Visit Ibiza Nightclubs: Your Ultimate Guide to the Island’s Hottest Spots appeared first on Geeky Traveller.

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If you run an eCommerce website, you may prefer to use a graphic as the call-to-action button for your subscription-style products or services. Below you can find a number of different subscription buttons suitable for use on a website’s landing page or the product page. Want some Buy Now button or Add to Cart button […]

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Lys234 is one of the residues present in class A β-lactamases that is under selective pressure due to antibiotic use. Located adjacent to proton shuttle residue Ser130, it is suggested to play a role in proton transfer during catalysis of the antibiotics. The mechanism underpinning how substitutions in this position modulate inhibitor efficiency and substrate specificity leading to drug resistance is unclear. The K234R substitution identified in several inhibitor-resistant β-lactamase variants is associated with decreased potency of the inhibitor clavulanic acid, which is used in combination with amoxicillin to overcome β-lactamase–mediated antibiotic resistance. Here we show that for CTX-M-14 β-lactamase, whereas Lys234 is required for hydrolysis of cephalosporins such as cefotaxime, either lysine or arginine is sufficient for hydrolysis of ampicillin. Further, by determining the acylation and deacylation rates for cefotaxime hydrolysis, we show that both rates are fast, and neither is rate-limiting. The K234R substitution causes a 1500-fold decrease in the cefotaxime acylation rate but a 5-fold increase in kcat for ampicillin, suggesting that the K234R enzyme is a good penicillinase but a poor cephalosporinase due to slow acylation. Structural results suggest that the slow acylation by the K234R enzyme is due to a conformational change in Ser130, and this change also leads to decreased inhibition potency of clavulanic acid. Because other inhibitor resistance mutations also act through changes at Ser130 and such changes drastically reduce cephalosporin but not penicillin hydrolysis, we suggest that clavulanic acid paired with an oxyimino-cephalosporin rather than penicillin would impede the evolution of resistance.

Sudip Ranjan Bhuia
Proc. Amer. Math. Soc. 152 (), 5249-5263.
Abstract, references and article information

Mammalian Asn-linked glycans are extensively processed as they transit the secretory pathway to generate diverse glycans on cell surface and secreted glycoproteins. Additional modification of the glycan core by α-1,6-fucose addition to the innermost GlcNAc residue (core fucosylation) is catalyzed by an α-1,6-fucosyltransferase (FUT8). The importance of core fucosylation can be seen in the complex pathological phenotypes of FUT8 null mice, which display defects in cellular signaling, development, and subsequent neonatal lethality. Elevated core fucosylation has also been identified in several human cancers. However, the structural basis for FUT8 substrate specificity remains unknown.Here, using various crystal structures of FUT8 in complex with a donor substrate analog, and with four distinct glycan acceptors, we identify the molecular basis for FUT8 specificity and activity. The ordering of three active site loops corresponds to an increased occupancy for bound GDP, suggesting an induced-fit folding of the donor-binding subsite. Structures of the various acceptor complexes were compared with kinetic data on FUT8 active site mutants and with specificity data from a library of glycan acceptors to reveal how binding site complementarity and steric hindrance can tune substrate affinity. The FUT8 structure was also compared with other known fucosyltransferases to identify conserved and divergent structural features for donor and acceptor recognition and catalysis. These data provide insights into the evolution of modular templates for donor and acceptor recognition among GT-B fold glycosyltransferases in the synthesis of diverse glycan structures in biological systems.

Broad-specificity glycoside hydrolases (GHs) contribute to plant biomass hydrolysis by degrading a diverse range of polysaccharides, making them useful catalysts for renewable energy and biocommodity production. Discovery of new GHs with improved kinetic parameters or more tolerant substrate-binding sites could increase the efficiency of renewable bioenergy production even further. GH5 has over 50 subfamilies exhibiting selectivities for reaction with β-(1,4)–linked oligo- and polysaccharides. Among these, subfamily 4 (GH5_4) contains numerous broad-selectivity endoglucanases that hydrolyze cellulose, xyloglucan, and mixed-linkage glucans. We previously surveyed the whole subfamily and found over 100 new broad-specificity endoglucanases, although the structural origins of broad specificity remained unclear. A mechanistic understanding of GH5_4 substrate specificity would help inform the best protein design strategies and the most appropriate industrial application of broad-specificity endoglucanases. Here we report structures of 10 new GH5_4 enzymes from cellulolytic microbes and characterize their substrate selectivity using normalized reducing sugar assays and MS. We found that GH5_4 enzymes have the highest catalytic efficiency for hydrolysis of xyloglucan, glucomannan, and soluble β-glucans, with opportunistic secondary reactions on cellulose, mannan, and xylan. The positions of key aromatic residues determine the overall reaction rate and breadth of substrate tolerance, and they contribute to differences in oligosaccharide cleavage patterns. Our new composite model identifies several critical structural features that confer broad specificity and may be readily engineered into existing industrial enzymes. We demonstrate that GH5_4 endoglucanases can have broad specificity without sacrificing high activity, making them a valuable addition to the biomass deconstruction toolset.

Arabinogalactan proteins (AGPs) are plant proteoglycans with functions in growth and development. However, these functions are largely unexplored, mainly because of the complexity of the sugar moieties. These carbohydrate sequences are generally analyzed with the aid of glycoside hydrolases. The exo-β-1,3-galactanase is a glycoside hydrolase from the basidiomycete Phanerochaete chrysosporium (Pc1,3Gal43A), which specifically cleaves AGPs. However, its structure is not known in relation to its mechanism bypassing side chains. In this study, we solved the apo and liganded structures of Pc1,3Gal43A, which reveal a glycoside hydrolase family 43 subfamily 24 (GH43_sub24) catalytic domain together with a carbohydrate-binding module family 35 (CBM35) binding domain. GH43_sub24 is known to lack the catalytic base Asp conserved among other GH43 subfamilies. Our structure in combination with kinetic analyses reveals that the tautomerized imidic acid group of Gln263 serves as the catalytic base residue instead. Pc1,3Gal43A has three subsites that continue from the bottom of the catalytic pocket to the solvent. Subsite −1 contains a space that can accommodate the C-6 methylol of Gal, enabling the enzyme to bypass the β-1,6–linked galactan side chains of AGPs. Furthermore, the galactan-binding domain in CBM35 has a different ligand interaction mechanism from other sugar-binding CBM35s, including those that bind galactomannan. Specifically, we noted a Gly → Trp substitution, which affects pyranose stacking, and an Asp → Asn substitution in the binding pocket, which recognizes β-linked rather than α-linked Gal residues. These findings should facilitate further structural analysis of AGPs and may also be helpful in engineering designer enzymes for efficient biomass utilization.

Litong Nie
Dec 1, 2020; 19:2015-2029
Research

The divalent anion sodium symporter (DASS) family (SLC13) plays critical roles in metabolic homeostasis, influencing many processes, including fatty acid synthesis, insulin resistance, and adiposity. DASS transporters catalyze the Na+-driven concentrative uptake of Krebs cycle intermediates and sulfate into cells; disrupting their function can protect against age-related metabolic diseases and can extend lifespan. An inward-facing crystal structure and an outward-facing model of a bacterial DASS family member, VcINDY from Vibrio cholerae, predict an elevator-like transport mechanism involving a large rigid body movement of the substrate-binding site. How substrate binding influences the conformational state of VcINDY is currently unknown. Here, we probe the interaction between substrate binding and protein conformation by monitoring substrate-induced solvent accessibility changes of broadly distributed positions in VcINDY using a site-specific alkylation strategy. Our findings reveal that accessibility to all positions tested is modulated by the presence of substrates, with the majority becoming less accessible in the presence of saturating concentrations of both Na+ and succinate. We also observe separable effects of Na+ and succinate binding at several positions suggesting distinct effects of the two substrates. Furthermore, accessibility changes to a solely succinate-sensitive position suggests that substrate binding is a low-affinity, ordered process. Mapping these accessibility changes onto the structures of VcINDY suggests that Na+ binding drives the transporter into an as-yet-unidentified conformational state, involving rearrangement of the substrate-binding site–associated re-entrant hairpin loops. These findings provide insight into the mechanism of VcINDY, which is currently the only structurally characterized representative of the entire DASS family.

The plasma membrane of a cell is characterized by an asymmetric distribution of lipid species across the exofacial and cytofacial aspects of the bilayer. Regulation of membrane asymmetry is a fundamental characteristic of membrane biology and is crucial for signal transduction, vesicle transport, and cell division. The type IV family of P-ATPases, or P4-ATPases, establishes membrane asymmetry by selection and transfer of a subset of membrane lipids from the lumenal or exofacial leaflet to the cytofacial aspect of the bilayer. It is unclear how P4-ATPases sort through the spectrum of membrane lipids to identify their desired substrate(s) and how the membrane environment modulates this activity. Therefore, we tested how the yeast plasma membrane P4-ATPase, Dnf2, responds to changes in membrane composition induced by perturbation of endogenous lipid biosynthetic pathways or exogenous application of lipid. The primary substrates of Dnf2 are glucosylceramide (GlcCer) and phosphatidylcholine (PC, or their lyso-lipid derivatives), and we find that these substrates compete with each other for transport. Acutely inhibiting sphingolipid synthesis using myriocin attenuates transport of exogenously applied GlcCer without perturbing PC transport. Deletion of genes controlling later steps of glycosphingolipid production also perturb GlcCer transport to a greater extent than PC transport. In contrast, perturbation of ergosterol biosynthesis reduces PC and GlcCer transport equivalently. Surprisingly, application of lipids that are poor transport substrates differentially affects PC and GlcCer transport by Dnf2, thus altering substrate preference. Our data indicate that Dnf2 exhibits exquisite sensitivity to the membrane composition, thus providing feedback onto the function of the P4-ATPases.

Insulin receptor substrate 2 (IRS2) is an essential adaptor that mediates signaling downstream of the insulin receptor and other receptor tyrosine kinases. Transduction through IRS2-dependent pathways is important for coordinating metabolic homeostasis, and dysregulation of IRS2 causes systemic insulin signaling defects. Despite the importance of maintaining proper IRS2 abundance, little is known about what factors mediate its protein stability. We conducted an unbiased proteomic screen to uncover novel substrates of the Anaphase Promoting Complex/Cyclosome (APC/C), a ubiquitin ligase that controls the abundance of key cell cycle regulators. We found that IRS2 levels are regulated by APC/C activity and that IRS2 is a direct APC/C target in G₁. Consistent with the APC/C's role in degrading cell cycle regulators, quantitative proteomic analysis of IRS2-null cells revealed a deficiency in proteins involved in cell cycle progression. We further show that cells lacking IRS2 display a weakened spindle assembly checkpoint in cells treated with microtubule inhibitors. Together, these findings reveal a new pathway for IRS2 turnover and indicate that IRS2 is a component of the cell cycle control system in addition to acting as an essential metabolic regulator.

Studies in the yeast Saccharomyces cerevisiae have helped define mechanisms underlying the activity of the ubiquitin–proteasome system (UPS), uncover the proteasome assembly pathway, and link the UPS to the maintenance of cellular homeostasis. However, the spectrum of UPS substrates is incompletely defined, even though multiple techniques—including MS—have been used. Therefore, we developed a substrate trapping proteomics workflow to identify previously unknown UPS substrates. We first generated a yeast strain with an epitope tagged proteasome subunit to which a proteasome inhibitor could be applied. Parallel experiments utilized inhibitor insensitive strains or strains lacking the tagged subunit. After affinity isolation, enriched proteins were resolved, in-gel digested, and analyzed by high resolution liquid chromatography-tandem MS. A total of 149 proteasome partners were identified, including all 33 proteasome subunits. When we next compared data between inhibitor sensitive and resistant cells, 27 proteasome partners were significantly enriched. Among these proteins were known UPS substrates and proteins that escort ubiquitinated substrates to the proteasome. We also detected Erg25 as a high-confidence partner. Erg25 is a methyl oxidase that converts dimethylzymosterol to zymosterol, a precursor of the plasma membrane sterol, ergosterol. Because Erg25 is a resident of the endoplasmic reticulum (ER) and had not previously been directly characterized as a UPS substrate, we asked whether Erg25 is a target of the ER associated degradation (ERAD) pathway, which most commonly mediates proteasome-dependent destruction of aberrant proteins. As anticipated, Erg25 was ubiquitinated and associated with stalled proteasomes. Further, Erg25 degradation depended on ERAD-associated ubiquitin ligases and was regulated by sterol synthesis. These data expand the cohort of lipid biosynthetic enzymes targeted for ERAD, highlight the role of the UPS in maintaining ER function, and provide a novel tool to uncover other UPS substrates via manipulations of our engineered strain.

Specific E3 ligases target tumor suppressors for degradation. Inhibition of such E3 ligases may be an important approach to cancer treatment. RNF146 is a RING domain and PARylation-dependent E3 ligase that functions as an activator of the β-catenin/Wnt and YAP/Hippo pathways by targeting the degradation of several tumor suppressors. Tankyrases 1 and 2 (TNKS1/2) are the only known poly-ADP-ribosyltransferases that require RNF146 to degrade their substrates. However, systematic identification of RNF146 substrates have not yet been performed. To uncover substrates of RNF146 that are targeted for degradation, we generated RNF146 knockout cells and TNKS1/2-double knockout cells and performed proteome profiling with label-free quantification as well as transcriptome analysis. We identified 160 potential substrates of RNF146, which included many known substrates of RNF146 and TNKS1/2 and 122 potential TNKS-independent substrates of RNF146. In addition, we validated OTU domain-containing protein 5 and Protein mono-ADP-ribosyltransferase PARP10 as TNKS1/2-independent substrates of RNF146 and SARDH as a novel substrate of TNKS1/2 and RNF146. Our study is the first proteome-wide analysis of potential RNF146 substrates. Together, these findings not only demonstrate that proteome profiling can be a useful general approach for the systemic identification of substrates of E3 ligases but also reveal new substrates of RNF146, which provides a resource for further functional studies.

Subsidies and Sustainable Agriculture: Mapping the Policy Landscape Research paper sysadmin 10 December 2019

Agricultural subsidies shape production and consumption patterns, with potentially significant effects on poverty, nutrition and other sustainability concerns. This paper maps the different types of support provided by governments to the agricultural sector, and highlights some of the complex political economy dynamics that underpin the relevant policies.

Summary

• Agricultural subsidies, a mainstay of government policy, have a large part in shaping production and consumption patterns, with potentially significant effects as regards poverty, food security, nutrition, and other sustainability concerns such as climate change, land use practices and biodiversity.
• There are multiple types of direct and indirect support provided by governments to various actors in the agricultural sector; and in terms of political economy, there are complex dynamics underpinning the policies that sustain these subsidies.
• Overall, subsidies targeting producers have the most significant effect on production, and the greater trade-distorting effect. These subsidies promote domestic production and discourage imports, leading to overproduction that is largely disposed of on the international market, with the help of export subsidies. This can tend to intensify negative environmental agricultural practices, such as cultivating marginal land, unsustainable types of intensification, or incentivizing excessive pesticide and fertilizer use.
• On the other hand, producer subsidies that are not tied to output of a specific commodity (i.e. delinked) have far fewer distorting impacts and could help to deliver sustainable outcomes. For example, this type of subsidies can require crop diversification or be linked to conservation of permanent grassland.
• Subsidies that enable transfers to consumers, for example through food stamp programmes, also serve to delink production from consumption, can foster healthier diets, can play an important role in delivering food accessibility and security among low-income groups, and can represent one of the less trade-distorting subsidies.
• If subsidies are to be reformed to help promote healthier diets and encourage more sustainable production, it is essential to understand not only the type and amount of support that key countries provide, but also the domestic dynamics that can shape such policies.
• While price support, input subsidies or investment aids remain the central pillars of programmes in large developing countries such as Brazil, China or India, other economies – notably including the EU and Japan – focus on direct payments, support for general services and set-aside schemes, as well as significant border protection. The US, for its part, has tended to focus on subsidized insurance schemes and food programmes for poorer consumers.
• If subsidies are to deliver policy objectives, their design and implementation should delink production from consumption. For example, consumer subsidies designed to deliver nutrition and food security, or payments for environmental services to enable more environmentally friendly production systems, could prove to be the most effective, least trade-distorting means of achieving more sustainable and equitable agricultural production.
• The political economy of food means that the removal of subsidies is often highly sensitive, and tends to be met with significant resistance. However, reform that delinks support from production through a gradual transition process could ultimately prove successful in delivering effective subsidy schemes.
• Effective subsidy schemes must by design be truly result- and performance-based, supported by robust and objective indicators. At the same time, engaging multiple actors along key commodity value chains – including leading importing and exporting countries, traders and transporters – could lead to the development of international, commodity-specific arrangements that are able to deliver effective nutrition and sustainability goals.

LONDON and TORONTO, Sept. 30, 2024 — Alphawave Semi, a global leader in high-speed connectivity and compute silicon for the world’s technology infrastructure, has unveiled the availability of the industry’s […]

The post Alphawave Semi and TSMC Collaborate on 3nm UCIe Die-to-Die Subsystem for AI and HPC appeared first on HPCwire.

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