HDAC6 controls autophagosome maturation essential for ubiquitin‐selective quality‐control autophagy

Autophagy is primarily considered a non‐selective degradation process induced by starvation. Nutrient‐independent basal autophagy, in contrast, imposes intracellular QC by selective disposal of aberrant protein aggregates and damaged organelles, a process critical for suppressing neurodegenerative diseases. The molecular mechanism that distinguishes these two fundamental autophagic responses, however, remains mysterious. Here, we identify the ubiquitin‐binding deacetylase, histone deacetylase‐6 (HDAC6), as a central component of basal autophagy that targets protein aggregates and damaged mitochondria. Surprisingly, HDAC6 is not required for autophagy activation; rather, it controls the fusion of autophagosomes to lysosomes. HDAC6 promotes autophagy by recruiting a cortactin‐dependent, actin‐remodelling machinery, which in turn assembles an F‐actin network that stimulates autophagosome–lysosome fusion and substrate degradation. Indeed, HDAC6 deficiency leads to autophagosome maturation failure, protein aggregate build‐up, and neurodegeneration. Remarkably, HDAC6 and F‐actin assembly are completely dispensable for starvation‐induced autophagy, uncovering the fundamental difference of these autophagic modes. Our study identifies HDAC6 and the actin cytoskeleton as critical components that define QC autophagy and uncovers a novel regulation of autophagy at the level of autophagosome–lysosome fusion.     

鲫鱼血清和皮肤粘液IgM的分离纯化及部分性质的鉴定

采用盐析法结合葡聚糖凝胶柱 ,分离纯化鲫鱼血清IgM ;然后制备兔抗鲫鱼血清IgM多克隆抗体 ,将其偶联到Sepharose 4B上制成亲和柱 ,用于分离纯化皮肤粘液IgM。结果表明 :33%～ 4 5 %硫酸铵溶液沉淀处理可以去除鲫鱼血清中除IgM外的很多杂蛋白 ,再经葡聚糖凝胶柱纯化 ,IgM纯度可达 80 %以上 ,其重链和轻链的分子量分别为 79和 2 5kDa ;以兔抗鲫鱼血清IgM多克隆抗体亲和柱分离皮肤粘液IgM ,分离效果良好 ,IgM重链的分子量为 88kDa ;Westernblot显示兔抗鲫鱼血清IgM多克隆抗体识别的是血清和皮肤粘液IgM的重链部分。用ELISA测定鲫鱼血清中IgM含量在一年中的变化 ,结果表明IgM在春夏季的含量高于秋冬季     

脂类物质抗菌的研究进展

研究发现人体内存在多种体内合成的抗菌物质,其中蛋白质类与糖类物质的研究报道较多。最近的研究发现脂类物质,包括饱和脂肪酸(月桂酸、棕榈酸)、单不饱和脂肪酸(棕榈烯酸、油酸)、多不饱和脂肪酸(二十碳五烯酸、二十二碳六烯酸、花生四烯酸)以及类固醇(维生素D、性类固醇)等也具有抗菌作用。脂类物质一方面可以通过调节多种炎性细胞和免疫细胞调动自身免疫系统发挥间接抗菌作用,另一方面也可以通过直接插入细菌细胞膜,影响其完整性发挥直接抗菌作用。本研究对各种脂类物质抗菌作用及其相关机制的进展作一综述。     

菠萝中蛋白酶及其他组份在品种间的差异分析

以湛江地区6个主要菠萝品种巴厘、卡因、台农17号、台农16号、粤脆、香水为对象,研究菠萝的组分在品种间的差异及其与菠萝蛋白酶的产率和酶活力之间的相关性,结果显示,不同的菠萝品种间,各组分的含量以及蛋白酶的得率及酶活有显著差异。菠萝果皮和果肉的不同生长部位的蛋白含量、酶活力及酶得率有显著差异。酶得率与蛋白及纤维素含量呈显著正相关;酶活力与糖酸比、得汁率、纤维素含量及Zn含量呈显著负相关,蛋白含量、纤维素含量、糖酸比、得汁率是衡量菠萝产酶品质的重要指标。     

Receptor-like cytokinin-binding protein(s) from barley leaves

Purified fractions of soluble proteins from barley leaves have been shown to contain specific binding sites fortrans-zeatin, a natural cytokinin. Such binding is very strong in vitro in concentrated solutions of some salts (ammonium sulfate or potassium phosphate) with optimum at pH 7–8 and temperature within the range 0–20°C. The cytokinin-binding sites have high affinity for zeatin (K_d1.5·10^–8 M) and low capacity corresponding to 1–1.5 pmol zeatin per milligram of initial soluble protein. Cytokinin binding is reversible; it is due to protein (or proteins) with molecular weight 40–45 kDa. This protein(s) does not bind³H-adenine and³H-abscisic acid. The ability of various compounds to displace³H-zeatin from its high-affinity binding sites is in strict accordance with their biological cytokinin activities. Other phytohormones as well as fusicoccin do not displace³H-zeatin from its binding sites. Specific zeatin binding is sensitive to heat, alkali, and pronase, but not to RNase treatment. The 150- to 200-fold purification of cytokinin-binding proteins was achieved by a combination of ammonium sulfate precipitation and Ultrogel AcA-54- and DEAE-cellulose chromatography. The zeatin-binding protein(s) from barley leaves is suggested to take part in cytokinin action in vivo.     

Selection and properties of mutant yeast <Emphasis Type="Italic">Pichia guilliermondii</Emphasis> strains resistant to chromium (VI)

Yeast Pichia guilliermondii strains L3 and L2, exposed to UV mutagenesis, produced over 80 mutants capable of growing on media containing 1.5 mM bichromate (Cr(VI)). The mutations making the strains resistant to Cr(VI) were dominant or semidominant. The mutants varied in Cr(VI) resistance, the degree of chromium accumulation in the cells (from 0.1 to 11.6 mg/g dry cells), and the degree of Cr(VI) reduction (from 50% to complete disappearance of bichromate from the culture liquid). Chromium accumulation in mutant cells depended on medium composition, Cr(VI) concentration, and the time of exposure to Cr(VI). The resistance to bichromate can be caused by various reasons: decrease in chromium absorption, altered ability to reduce Cr(VI), or damage of sulfate transport mechanisms.Translated from Prikladnaya Biokhimiya i Mikrobiologiya, Vol. 41, No. 2, 2005, pp. 204–209.Original Russian Text Copyright © 2005 by Babyak, Ksheminskaya, Gonchar, Yanovich, Fedorovich.     

灌浆期高温对冬小麦籽粒氨基酸含量和组成的影响

以强筋小麦品种郑麦366和中筋小麦品种洛旱2号为材料,采用盆栽和人工气候室模拟的方法,研究了花后不同时段(灌浆前期即花后5 d,灌浆中期即花后15 d)和持续时间(处理2 d和4 d)的高温胁迫对籽粒氨基酸含量和积累量(以单粒中氨基酸含量表示)的影响。结果显示:(1)高温胁迫显著增加了小麦籽粒中必需氨基酸(EAA)、非必需氨基酸(NAA)和总氨基酸(TAA)含量,但降低了其积累量和EAA/TAA,2品种表现一致;(2)灌浆前期高温胁迫对2品种籽粒氨基酸含量的影响大于灌浆中期,而对氨基酸积累量的影响则相反;(3)高温持续时间对籽粒氨基酸含量的影响在2品种间存在差异,洛旱2号籽粒赖氨酸、EAA、NAA和TAA含量均随高温持续时间的延长显著增加,而郑麦366籽粒中上述指标仅在高温胁迫4 d下较对照增加显著;(4)从受高温胁迫的影响看,籽粒EAA/TAA对高温时段更敏感,而氨基酸含量表现为更易受高温持续时间的影响;(5)高温时段与持续时间的互作效应体现在:灌浆前期籽粒氨基酸积累量2品种均以高温胁迫4 d的影响最大,而灌浆中期则均以高温2 d的降幅最大。上述结果表明,小麦籽粒氨基酸及其组分对高温胁迫的响应不仅在品种间存在差异,且受高温时段和持续时间的影响。     

急性冷应激对牦牛乳腺上皮细胞 HSP70 mRNA 表达的影响

本文主要研究了急性冷应激对牦牛乳腺上皮细胞热休克蛋白７０ （Heat stress protein,ＨＳＰ７０） 表达量的影响。采用实时荧光定量ＰＣＲ 技术,以急性冷刺激１０℃ 为典型研究环境,分析了ＨＳＰ７０ ｍＲＮＡ 表达的变化规律。结果显示,乳腺上皮细胞在１０℃分别冷处理２ ｈ、４ ｈ、６ ｈ 和８ ｈ,其ＨＳＰ７０ ｍＲＮＡ 的表达量变化均不显著（Ｐ ＞０. ０５）;分别在１０℃冷处理２ ｈ、４ ｈ、６ ｈ 和８ ｈ,再复温培养４ ｈ,ＨＳＰ７０ ｍＲＮＡ 的表达量均极显著增加（Ｐ ＜０. ０１）,于６ ｈ 达到峰值;在１０℃先冷处理４ ｈ,然后分别复温２ ｈ、４ ｈ、６ ｈ 和８ ｈ,ＨＳＰ７０ ｍＲＮＡ 的表达量亦均显著增加（Ｐ ＜０. ０１）,并于４ ｈ 达到峰值。结论：急性冷应激诱导牦牛乳腺上皮细胞ＨＳＰ７０ 表达量的增加不是发生在冷处理过程中,而是发生在复温过程中,并且在一定范围内随冷处理时间的增长表达量增高。     

Spatio-temporal expression of Prospero is finely tuned to allow the correct development and function of the nervous system in Drosophila melanogaster

Adaptive animal behaviors depend upon the precise development of the nervous system that underlies them. In Drosophila melanogaster, the pan-neural prospero gene (pros), is involved in various aspects of neurogenesis including cell cycle control, axonal outgrowth, neuronal and glial cell differentiation. As these results have been generally obtained with null pros mutants inducing embryonic lethality, the role of pros during later development remains poorly known. Using several pros-Voila (prosV) alleles, that induce multiple developmental and behavioral anomalies in the larva and in adult, we explored the relationship between these phenotypes and the variation of pros expression in 5 different neural regions during pre-imaginal development. We found that the quantity of pros mRNA spliced variants and of Pros protein varied between these alleles in a tissue-specific and developmental way. Moreover, in prosV1 and prosV13 alleles, the respective decrease or increase of pros expression, affected (i) neuronal and glial cell composition, (ii) cell proliferation and death and (iii) axonal-dendritic outgrowth in a stage and cellular context dependant way. The various phenotypic consequences induced during development, related to more or less subtle differences in gene expression, indicate that Pros level needs a precise and specific adjustment in each neural organ to allow its proper function.