Cryo-EM structure of DNA-bound Smc5/6 reveals DNA clamping enabled by multi-subunit conformational changes

Structural maintenance of chromosomes (SMC) complexes are essential for chromatin organization and functions throughout the cell cycle. The cohesin and condensin SMCs fold and tether DNA, while Smc5/6 directly promotes DNA replication and repair. The functions of SMCs rely on their abilities to engage DNA, but how Smc5/6 binds and translocates on DNA remains largely unknown. Here, we present a 3.8 Å cryogenic electron microscopy (cryo-EM) structure of DNA-bound Saccharomyces cerevisiae Smc5/6 complex containing five of its core subunits, including Smc5, Smc6, and the Nse1-3-4 subcomplex. Intricate interactions among these subunits support the formation of a clamp that encircles the DNA double helix. The positively charged inner surface of the clamp contacts DNA in a nonsequence-specific manner involving numerous DNA binding residues from four subunits. The DNA duplex is held up by Smc5 and 6 head regions and positioned between their coiled-coil arm regions, reflecting an engaged-head and open-arm configuration. The Nse3 subunit secures the DNA from above, while the hook-shaped Nse4 kleisin forms a scaffold connecting DNA and all other subunits. The Smc5/6 DNA clamp shares similarities with DNA-clamps formed by other SMCs but also exhibits differences that reflect its unique functions. Mapping cross-linking mass spectrometry data derived from DNA-free Smc5/6 to the DNA-bound Smc5/6 structure identifies multi-subunit conformational changes that enable DNA capture. Finally, mutational data from cells reveal distinct DNA binding contributions from each subunit to Smc5/6 chromatin association and cell fitness. In summary, our integrative study illuminates how a unique SMC complex engages DNA in supporting genome regulation.

Structural maintenance of chromosomes (SMC) complexes are essential genome regulators in both prokaryotes and eukaryotes. In eukaryotes, the cohesin and condensin SMC complexes organize DNA, while the Smc5/6 complex (referred to as Smc5/6) directly regulates DNA replication and repair (1). At the structural level, SMC complexes share similarities while possessing unique attributes (1). Each complex contains a pair of SMC subunits and a set of non-SMC subunits. The SMC subunits define the tripartite filamentous architecture of the complex: their approximal 50-nm long coiled coil arm region connects their dimerized hinge and adenosine triphosphatase (ATPase) head regions (1). A non-SMC kleisin subunit uses its N- and C-terminal domains to link the head of one SMC to the head-proximal arm region (neck) of another SMC, forming a trimeric SMC-kleisin structure. In cohesin and condensin, two large U-shaped HEAT (Huntington, elongation factor 3, PR65/A, TOR) repeat HAWK (HEAT proteins associated with kleisins) subunits attach to the middle region of the kleisin. By contrast, the Smc5/6 kleisin (Nse4) binds to smaller WH (winged helix)-containing KITE (kleisin interacting tandem WH elements) subunits (Nse1 and Nse3) (2).SMC-mediated functions depend on interactions with DNA. Recent cryogenic electron microscopy (cryo-EM) structures of DNA-bound cohesin and condensin revealed that their HAWK subunits and the SMC head-neck regions form a clamp to enclose a single DNA double helix (3–7). DNA clamping can be critical for cohesin and condensin to extrude DNA loops for chromatin folding (5, 7–9). DNA loop extrusion additionally requires arm bending at a region called the elbow, which is found in both cohesin and condensin (5, 7–9). By contrast, a lack of arm bending in Smc5/6 was suggested by negative stain EM and cross-linking mass spectrometry (CLMS) data (10–14). Since Smc5/6 does not contain HAWK proteins nor shows arm-bending, it has remained unclear how Smc5/6 engages DNA to accomplish its multiple functions.Here we address the molecular mechanisms by which this unique SMC complex binds DNA using an integrative approach, coupling a cryo-EM-based structural characterization with CLMS analyses and functional investigation. Our atomic structure of a DNA-bound Saccharomyces cerevisiae Smc5/6 complex reveals that the head-neck Smc5-6 regions and the Nse1-3-4 subcomplex together form a clamp entrapping the DNA helix. The structure further reveals protein subunit folds and association, as well as how the subunits collaborate to entrap DNA. Comparison of CLMS analyses of DNA-free Smc5/6 with the structure of the DNA-bound Smc5/6 unveils large scale, multi-subunit conformational changes that enable Smc5/6 to encircle DNA. Finally, our mutational data suggest distinct contributions from each of the DNA binding regions to Smc5/6 chromatin association and cellular fitness. Comparison of our findings with those of other SMCs reveals that diverse SMC complexes use a similar DNA clamping strategy despite structural differences, and that Smc5/6 possesses unique features distinct from cohesin, condensin, and prokaryotic SMCs. Our work lays the foundation for an in-depth understanding of how Smc5/6 fulfills unique roles in genome protection.     

Dasatinib and interferon alpha synergistically induce pyroptosis-like cell death in philadelphia chromosome positive acute lymphoblastic leukemia

Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL) is a high-risk disease subtype with a dismal prognosis. Inhibiting BCR-ABL kinase alone is insufficient to eradicate Ph+ALL clones, and alternative BCR-ABL-dependent and -independent pathways need to be targeted as an effective strategy. Our study revealed that the combination of dasatinib and interferon-α showed synergistic activity against Ph+ALL, inducing mitochondrial dysfunction and causing necrosis-like cell lysis. Mechanistic studies showed that the induced cell death was caspase-3-independent. Canonical necroptosis signals, such as RIP1 and MLKL, were not activated; instead, the pyroptosis executor Gasdermin D was upregulated expression and activated. The expression levels of extracellular ATP and IL-1β were also upregulated, both of which are markers of pyroptotic cell death. In a murine Ph+ALL model, the dual drug treatment prolonged the survival of tumor-bearing mice. More importantly, we incorporated the dual drugs to maintenance therapy in 39 patients who were unfit for allogeneic stem cell transplantation (allo-HSCT). The median follow-up was 28.5 months, the 4-year disease-free survival and overall survival rates were 52.2% and 65.2%, respectively. Our data suggest that the combination of dasatinib and interferon-α has potential synergistic activity against Ph+ALL and shows promise as a maintenance therapy for Ph+ALL patients who are unfit for allo-HSCT.     

Association of body mass index with risk of prediabetes in Chinese adults: A population‐based cohort study

Aims/IntroductionOverweight and obesity in adults are strongly associated with an increased risk of prediabetes, and this study set out to gain a better understanding of the optimal body mass index (BMI) range for assessing the risk of prediabetes in the Chinese population.Materials and MethodsThe cohort study included 100,309 Chinese adults who underwent health screening. Participants were divided into six groups based on the cut‐off point for BMI recommended by the World Health Organization (underweight: <18.5 kg/m², normal‐weight: 18.5–24.9 kg/m², pre‐obese: 25.0–29.9 kg/m², obese class I: 30.0–34.9 kg/m², obese class II: 35.0–39.9 kg/m², and obese class III ≥40 kg/m²). The association of BMI with prediabetes and the shape of the correlation were modeled using multivariate Cox regression and restricted cubic spline regression, respectively.ResultsIn the multivariate Cox regression model, with normal weight as the control group, underweight people had a lower risk of developing prediabetes, whereas obese and pre‐obese people had a higher risk of prediabetes. Additionally, in the restricted cubic spline model, we found that the association of BMI with prediabetes follows a positive dose–response relationship, but does not conform to the pattern of obesity paradox. Among the general population in China, a BMI of 23.03 kg/m² might be a potential intervention threshold for prediabetes.ConclusionsThe national cohort study found that the association of BMI with prediabetes follows a positive dose–response relationship, rather than a pattern of obesity paradox. For Chinese people with normal weight, more attention should be paid to glucose metabolism when BMI exceeds 23.03 kg/m².     

传染病新特点与口岸卫生检疫应对

目的提高国境口岸应对传染病的新特点的能力。方法分析传染病新近出现新的特点和趋势,提出国境口岸卫生检疫的应对措施。结果应对传染病出现全球化、卷土重来的新特点,口岸卫生检疫工作任重道远,加强国际、国内、区域间及人医与兽医的合作,调整卫生检疫模式及内容,建立有效的防控体系。结论在传染病全球化和增多的情况下,建立有效的卫生检疫模式。     

NT-3、NT-4和BDNF在面神经逆行转运的动力学比较

目的　研究神经营养素 3(NT- 3)、神经营养素 4(NT- 4)和脑源性神经营养因子 (BDNF)在面神经中的逆行转运动力学特点。方法　将新西兰兔一侧面神经干横切断后置入硅胶再生室 ,再将 1 2 5I标记的 NT- 3或NT- 4或 BDNF或人血清白蛋白 (HSA) (10μl/只 ,约 3.7MBq)注入再生室 ,在注药后于不同时刻取兔面神经干和脑干面神经运动核 ,测定其摄取率。利用 3P87动力学处理程序分析计算各标记物在面神经的动力学参数。结果　面神经逆行转运神经营养素的转运量 :NT- 3>BDNF>NT- 4(P <0 .0 5 ) ,转运速率 :NT- 4>NT- 3>BDNF (P <0 .0 5 )。结论　研究结果提供了神经营养素在面神经逆行转运的动力学特点。     

Arthroscopic Intra‐Articular Repair of the Torn Subscapularis Tendon with Single Anterior Portal,Single Suture Anchor,and X‐Shaped Fixation Technique

Subscapular tendon plays an important role in shoulder joint function. With the advance of magnetic resonance imaging technology and the popularization of arthroscopic shoulder surgery, subscapularis tears have been increasingly detected. However, reduction and fixation of subscapular tendon tears appears to be technically challenging. This study aims to describe an arthroscopic intra‐articular X‐shaped fixation technique: a procedure of subscapularis tendon repair performed with the aid of a suture passer using only a single anterior portal and a single suture anchor. By incorporating the advantages of a single anterior working portal for anchor placement and tear repair, this technique provides an easier way to use suture lasso and make knots in a limited working space, and the whole procedure is minimally invasive with a short learning curve. This technique has been applied in patients with subscapularis tears involving no intraoperative or postoperative complications. Our technology offers a valuable new treatment option for subscapularis tears.     

血清可溶性白细胞介素2受体和T细胞亚群在单纯性肾病综合征患儿发病中作用的探讨

本文对26例儿童单纯性肾病综合征患者血清可溶性白细胞介素2受体（SIL-2R）及T细胞亚群进行研究。结果表明肾病活动期SIL-2R值升高（均值为895±245U／ml），CD_4~+／CD_8~+比值降低（P＜0.01），而肾病缓解期SIL-2R（均值为494±127U／ml）及CD_4~+／CD_8~+比值恢复正常，将SIL-2R值与CD_4~+／CD_8~+比值进行相关分析，发现两者呈负相关（r=-0.61，P＜0.05）。提示SIL-2R水平的升高及T细胞亚群的紊乱在儿童单纯性肾病发病机制中起到重要的作用。     

箝式取栓治疗颅内大血管栓塞的体外模型

临床研究显示,与传统支架取栓(CSRT)比较,箝式取栓(TCET)可降低术中栓子碎裂逃逸概率,提高取栓成功率[1]。本实验旨在探讨TCET取栓机制并与CSRT比较取栓术中栓子逃逸情况。     

Dextromethorphan Attenuated Inflammation and Combined Opioid Use in Humans Undergoing Methadone Maintenance Treatment

Journal of Neuroimmune Pharmacology - Recent studies show that proinflammatory cytokines might be related to the development of opioid dependence (physiological, psychological, or both). In a...