调控细胞活动不可或缺的重要分子——F-box蛋白

泛素连接酶作为一种翻译后效应器,对细胞生命活动的正常运行至关重要。而泛素连接酶SCF(Skp1-Cullin-F-box protein)复合体重要组件—F-box蛋白的主要作用是对靶蛋白的特异性识别。作为许多生理病理过程的效应分子,它广泛存在于真核生物,参与了众多的细胞机制,其对底物的特异性识别是蛋白元件在特定时空功能终止的重要基础。对F-box蛋白的深入研究必将增强人们对细胞生命活动调控机制和疾病机理的理解。本文拟就F-box蛋白的结构、功能及其与疾病发生的研究进展做一综述。     

Autophagy Reduces Neuronal Damage and Promotes Locomotor Recovery via Inhibition of Apoptosis After Spinal Cord Injury in Rats

Autophagy is an intracellular catabolic mechanism that maintains the balance of proteins, lipids and aging organelles. 3-Methyladenine (3-MA) is a selective inhibitor of autophagy, whereas rapamycin, an antifungal agent, is a specific inducer of autophagy, inhibiting the protein mammalian target of rapamycin. In the present study, we examined the role of autophagy, inhibited by 3-MA and enhanced by rapamycin, in a model of acute spinal cord injury in rats. We found that rapamycin could significantly increase the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 at the injury site. At the same time, the number of neurons and astrocytes with LC3 positive in the spinal cord was upregulated with time. In addition, administration of rapamycin produced an increase in the Basso, Beattie and Bresnahan scores of injured rats, indicating high recovery of locomotor function. Furthermore, expression of the proteins Bcl-2 and Bax was upregulated and downregulated, respectively. By contrast, the results for rats treated with 3-MA, which inhibits autophagy, were the opposite of those seen with the rapamycin-treated rats. These results show that induction of autophagy can produce neuroprotective effects in acute spinal cord injury in rats via inhibition of apoptosis.     

Structure and function of flavivirus NS5 methyltransferase

The plus-strand RNA genome of flavivirus contains a 5' terminal cap 1 structure (m7GpppAmG). The flaviviruses encode one methyltransferase, located at the N-terminal portion of the NS5 protein, to catalyze both guanine N-7 and ribose 2'-OH methylations during viral cap formation. Representative flavivirus methyltransferases from dengue, yellow fever, and West Nile virus (WNV) sequentially generate GpppA-->m7GpppA-->m7GpppAm. The 2'-O methylation can be uncoupled from the N-7 methylation, since m7GpppA-RNA can be readily methylated to m7GpppAm-RNA. Despite exhibiting two distinct methylation activities, the crystal structure of WNV methyltransferase at 2.8 A resolution showed a single binding site for S-adenosyl-L-methionine (SAM), the methyl donor. Therefore, substrate GpppA-RNA should be repositioned to accept the N-7 and 2'-O methyl groups from SAM during the sequential reactions. Electrostatic analysis of the WNV methyltransferase structure showed that, adjacent to the SAM-binding pocket, is a highly positively charged surface that could serve as an RNA binding site during cap methylations. Biochemical and mutagenesis analyses show that the N-7 and 2'-O cap methylations require distinct buffer conditions and different side chains within the K61-D146-K182-E218 motif, suggesting that the two reactions use different mechanisms. In the context of complete virus, defects in both methylations are lethal to WNV; however, viruses defective solely in 2'-O methylation are attenuated and can protect mice from later wild-type WNV challenge. The results demonstrate that the N-7 methylation activity is essential for the WNV life cycle and, thus, methyltransferase represents a novel target for flavivirus therapy.     

紫花苜蓿对铜胁迫生理响应的傅里叶变换红外光谱法研究

为阐明紫花苜蓿(Medicago sativa L.)对铜胁迫的耐性机理,采用准确度好、分辨率高和简便快捷的傅里叶变换红外光谱法(FTIR)研究在不同铜浓度(0、1、5、20、100 mol/L)处理时紫花苜蓿根、茎、叶化学组分的变化。结果表明:随着铜处理浓度的增加,紫花苜蓿根、茎、叶生物量变化不大。其根组织在2924 cm~(-1)处峰高处呈现出先下降后上升的趋势,反映了在低铜(5 mol/L)处理条件下紫花苜蓿分泌的有机酸不断螯合Cu,造成羧酸0-H的减少,但随着Cu含量的升高,其羧酸螯合力变弱,有机酸含量渐渐升高;根组织在1381 cm~(-1)处峰高先下降后上升,反映了含油脂化合物含量先下降后升高。可能与植物在细胞壁结构上增强抗逆性有一定关系,即低Cu处理下细胞壁可能通过提高阳离子交换能力(CEC),增强了耐Cu性;茎组织在2924,1643,1381,1064 cm~(-1)等处峰高无明显变化;叶组织所有峰值在低浓度(5 mol/L)Cu处理下变化不明显,高浓度(5 moL/L)Cu处理下所有峰值先升后降,随着这可能与可溶性糖及可溶性蛋白质等物质含量都呈现先升后降的趋势有关。这表明紫花苜蓿通过根部有机酸含量的变化和提高细胞壁阳离子交换能力,将吸收的Cu大部分积累在根部,阻止Cu向地上部分运输,有效地保护了植物地上部分组织。     

缙云山细枝柃种群性比及空间分布

缙云山细枝柃（Eurya loquaiana）种群包含一些性别变异个体,表现出"亚雌雄异株"的特点。为分析该种群的性别格局,了解柃木属植物性别系统的进化及其适应机制,在缙云山细枝柃分布的不同生境中设置8个20 m×20 m长期观测样地,分析性比变化及其与环境、年龄结构的关系,同时设置1个20 m×60 m样地,分析植株的空间分布格局及性别间在空间上的关联。结果显示：（1）缙云山细枝柃种群总体上性别偏雌,雌雄比为1.43,性别变异株比例达14.42%。（2）各样地中雌雄比和性别变异株比例差异较大,性别变异株比例与干扰强度呈较强正相关而与海拔高度和郁闭度呈较强负相关,显示变异株的出现可能与人为干扰有一定联系;雌雄比与样地中细枝柃平均基径显著正相关,可能随着林龄的增加,雌雄比有增加的趋势。（3）细枝柃个体在空间上整体呈聚集分布,雌株、雄株及性别变异株主要呈随机分布,在部分尺度偏向于聚集分布。（4）雌株和雄株对环境异质性的响应相似,在空间分布上有一定的正相关。上述结果反应了缙云山细枝柃种群的性别格局及其生态适应机制。     

Natural diterpenoid compound elevates expression of Bim protein, which interacts with antiapoptotic protein Bcl-2, converting it to proapoptotic Bax-like molecule

Overwhelming evidence indicates that Bax and Bak are indispensable for mediating cytochrome c release from mitochondria during apoptosis. Here we report a Bax/Bak-independent mechanism of cytochrome c release and apoptosis. We identified a natural diterpenoid compound that induced apoptosis in bax/bak double knock-out murine embryonic fibroblasts and substantially reduced the tumor growth from these cells implanted in mice. Treatment with the compound significantly increased expression of Bim, which migrated to mitochondria, altering the conformation of and forming oligomers with resident Bcl-2 to induce cytochrome c release and caspase activation. Importantly, purified Bim and Bcl-2 proteins cooperated to permeabilize a model mitochondrial outer membrane; this was accompanied by oligomerization of these proteins and deep embedding of Bcl-2 in the membrane. Therefore, the diterpenoid compound induces a structural and functional conversion of Bcl-2 through Bim to permeabilize the mitochondrial outer membrane, thereby inducing apoptosis independently of Bax and Bak. Because Bcl-2 family proteins play important roles in cancer development and relapse, this novel cell death mechanism can be explored for developing more effective anticancer therapeutics.