Crystal structure and functional analysis of MiD49, a receptor for the mitochondrial fission protein Drp1

Mitochondrial fission requires recruitment of dynamin‐related protein 1 (Drp1) to the mitochondrial surface, where assembly leads to activation of its GTP‐dependent scission function. MiD49 and MiD51 are two receptors on the mitochondrial outer membrane that can recruit Drp1 to facilitate mitochondrial fission. Structural studies indicated that MiD51 has a variant nucleotidyl transferase fold that binds an ADP co‐factor essential for activation of Drp1 function. MiD49 shares sequence homology with MiD51 and regulates Drp1 function. However, it is unknown if MiD49 binds an analogous co‐factor. Because MiD49 does not readily crystallize, we used structural predictions and biochemical screening to identify a surface entropy reduction mutant that facilitated crystallization. Using molecular replacement, we determined the atomic structure of MiD49 to 2.4 Å. Like MiD51, MiD49 contains a nucleotidyl transferase domain; however, the electron density provides no evidence for a small‐molecule ligand. Structural changes in the putative nucleotide‐binding pocket make MiD49 incompatible with an extended ligand like ADP, and critical nucleotide‐binding residues found in MiD51 are not conserved. MiD49 contains a surface loop that physically interacts with Drp1 and is necessary for Drp1 recruitment to the mitochondrial surface. Our results suggest a structural basis for the differential regulation of MiD51‐ versus MiD49‐mediated fission.     

秸秆全量还田对稻田土壤溶解有机碳含量和水稻产量的影响

以南粳44为供试材料,在粘土和砂土土壤中,设置麦秸秆不还田和全量还田(6000 kg·hm^-2)及3种施氮量(0、225、300 kg·hm^-2)试验,研究了麦秸秆全量还田的腐解率和有机碳释放量动态变化,及其对稻田0～45 cm土壤溶解有机碳(DOC)含量和水稻产量的影响.结果表明： 麦秸秆还田的前期(0～30 d)其腐解率和有机碳释放量最高,腐解率为35.0%(粘土)和31.7%(砂土),有机碳释放率为34.1%(粘土)和33.1%(砂土);30 d后两者均减小.施用氮肥可显著促进秸秆腐解和有机碳释放量,粘土中麦秸秆腐解率和有机碳释放量明显大于砂土.麦秸秆还田后土壤DOC含量逐渐增加,至25 d达最大值,粘土和砂土分别为60.18和56.62 mg·L^-1,此后逐渐减小并趋于稳定.麦秸秆还田处理15 cm处土壤DOC含量显著高于未还田处理,但两者在30和45 cm处土壤DOC含量差异不显著,说明秸秆还田主要增加了稻田0～15 cm土层DOC含量.与不施氮处理相比,施氮处理土壤DOC含量降低,2种施氮处理间差异不显著.秸秆还田减少了水稻前期分蘖发生量,显著降低了有效穗数,增加了穗粒数、结实率和千粒重,显著提高了水稻产量.     

烟草叶片对不同光质短期应答的代谢轮廓

在烟草上部叶片生理成熟期给以白、紫外A、蓝、绿、黄、红6种LED灯光处理,研究了烟草叶片对不同光质的短期应答.结果表明: 采用非靶标检测方法共检测到68种GC/MS(气相色谱/质谱)稳定的代谢产物,在PLS-DA得分图中,6种光处理的样品被明显分开;有61种在质谱库中得到鉴定,其中45种代谢物质含量在不同光处理间存在显著性差异,主要为有机酸类、糖类、TCA循环中间产物、氨基酸类等初级代谢产物;采用差异性代谢物的热力图及聚类分析将其分为5类,同时也将6个处理明显分开,其中红、蓝光处理差异最为明显,除B类物质外,其他4类均以红光处理高于蓝光处理.采用靶标性检测方法测定了3种生物碱、5种多酚及茄尼醇的含量,4种(加上非靶标测定到的烟碱)生物碱在不同光质处理间的变化趋势一致,红光和黄光明显促进了生物碱积累;莰菲醇基芸香苷和芸香苷在不同处理之间的变化趋势一致,以蓝光处理最低、黄光最高,光质对其他3种多酚的影响各异;黄光明显抑制了茄尼醇的积累,蓝光处理的叶片中茄尼醇含量最高.说明光质变化对烟草脂肪酸代谢、糖代谢、生物碱代谢、氨基酸代谢、TCA循环和莽草酸途径等多条代谢途径均有显著的影响.     

秦岭滑蜥排泄系统组织形态学观察

运用大体解剖和组织切片技术对秦岭滑蜥Scincella tsinlingensis排泄系统进行了组织形态学观察。结果显示:秦岭滑蜥的排泄系统包括肾脏、输尿管、膀胱和泄殖腔。肾脏由被膜与实质构成,实质包括许多泌尿小管与少量结缔组织。泌尿小管包括肾单位与集合管,肾单位的数量较两栖类有了明显增加。结缔组织中分布有少量弹性纤维、网状纤维和大量胶原纤维。输尿管由许多分支的集尿管汇聚形成,包括黏膜与外膜,黏膜由单层柱状上皮过渡为多层鳞状细胞,固有层零散分布有浆细胞,结缔组织中分布有胶原纤维与少量弹性纤维。膀胱由黏膜层、肌层与外膜构成,黏膜上皮为变移上皮,固有层分布有少量的浆细胞。消化道、生殖道和输尿管末端汇聚于泄殖腔,泄殖腔壁由黏膜、肌层和外膜构成,黏膜上皮分布有黏液性细胞和少量浆细胞。秦岭滑蜥的排泄系统与其他卵胎生蜥蜴无明显差别。     

Combination of inflammation-related cytokines promotes long-term muscle stem cell expansion

Muscle stem cells (MuSCs, satellite cells) are the major contributor to muscle regeneration. Like most adult stem cells, long-term expansion of MuSCs in vitro is difficult. The in vivo muscle regeneration abilities of MuSCs are quickly lost after culturing in vitro, which prevents the potential applications of MuSCs in cell-based therapies. Here, we establish a system to serially expand MuSCs in vitro for over 20 passages by mimicking the endogenous microenvironment. We identified that the combination of four pro-inflammatory cytokines, IL-1α, IL-13, TNF-α, and IFN-γ, secreted by T cells was able to stimulate MuSC proliferation in vivo upon injury and promote serial expansion of MuSCs in vitro. The expanded MuSCs can replenish the endogenous stem cell pool and are capable of repairing multiple rounds of muscle injuries in vivo after a single transplantation. The establishment of the in vitro system provides us a powerful method to expand functional MuSCs to repair muscle injuries.     

Angiomotin binding-induced activation of Merlin/NF2 in the Hippo pathway

The tumor suppressor Merlin/NF2 functions upstream of the core Hippo pathway kinases Lats1/2 and Mst1/2, as well as the nuclear E3 ubiquitin ligase CRL4^DCAF1. Numerous mutations of Merlin have been identified in Neurofibromatosis type 2 and other cancer patients. Despite more than two decades of research, the upstream regulator of Merlin in the Hippo pathway remains unknown. Here we show by high-resolution crystal structures that the Lats1/2-binding site on the Merlin FERM domain is physically blocked by Merlin''s auto-inhibitory tail. Angiomotin binding releases the auto-inhibition and promotes Merlin''s binding to Lats1/2. Phosphorylation of Ser518 outside the Merlin''s auto-inhibitory tail does not obviously alter Merlin''s conformation, but instead prevents angiomotin from binding and thus inhibits Hippo pathway kinase activation. Cancer-causing mutations clustered in the angiomotin-binding domain impair angiomotin-mediated Merlin activation. Our findings reveal that angiomotin and Merlin respectively interface cortical actin filaments and core kinases in Hippo signaling, and allow construction of a complete Hippo signaling pathway.     

转BT+ CPTI基因抗虫棉的种植对土壤根际反硝化菌丰度和多样性影响

摘要:【目的】为评价转基因棉花的种植对根际土壤反硝化细菌丰度和多样性的影响,【方法】以转Bt + CpTI 基因抗虫棉(SGK321)及非转基因亲本棉花石远321(SY321)根际土壤反硝化细菌为研究对象,分别在种植前和棉花的不同生长时期(蕾期、花期、铃期和絮期)取样。采用实时荧光定量PCR方法和末端标记限制性片段多态性(T-RFLP)技术对土壤中反硝化细菌的功能基因nosZ基因进行定量和多样性分析。【结果】结果发现随棉花不同生长时期两种棉花根际土壤中的反硝化细菌丰度均发生显著变化,但变化趋势不同。转基因棉花根际土壤反硝化细菌从种植前的3.12×106 copies/g dry soil一直增加到铃期的2.81×107 copies/g dry soil,增加了8倍。非转基因棉花根际土壤反硝化细菌丰度变化受生长周期影响更为显著,表现为蕾期增加,花期降低,铃期又增加的趋势。典范相关及部分典范相关分析反硝化细菌多样性结果表明其多样性受环境因素pH、硝酸根浓度以及棉花生长时期(蕾期和花期)影响最为显著,但此外棉花品种也起到了非常重要的作用,【结论】反硝化细菌的丰度和多样性既受棉花生长时期的影响,同时也受棉花品种的影响,转基因棉花通过调节根际土壤中的pH 和硝酸根浓度,来影响其多样性和丰度。转Bt + CpTI基因抗虫棉的种植增加了土壤pH,从而导致根际土壤反硝化细菌的多样性和丰度增加。