Recruitment of Nox4 to a Plasma Membrane Scaffold Is Required for Localized Reactive Oxygen Species Generation and Sustained Src Activation in Response to Insulin-like Growth Factor-I

Nox4-derived ROS is increased in response to hyperglycemia and is required for IGF-I-stimulated Src activation. This study was undertaken to determine the mechanism by which Nox4 mediates sustained Src activation. IGF-I stimulated sustained Src activation, which occurred primarily on the SHPS-1 scaffold protein. In vitro oxidation experiments indicated that Nox4-derived ROS was able to oxidize Src when they are in close proximity, and Src oxidation leads to its activation. Therefore we hypothesized that Nox4 recruitment to the plasma membrane scaffold SHPS-1 allowed localized ROS generation to mediate sustained Src oxidation and activation. To determine the mechanism of Nox4 recruitment, we analyzed the role of Grb2, a component of the SHPS-1 signaling complex. We determined that Nox4 Tyr-491 was phosphorylated after IGF-I stimulation and was responsible for Nox4 binding to the SH2 domain of Grb2. Overexpression of a Nox4 mutant, Y491F, prevented Nox4/Grb2 association. Importantly, it also prevented Nox4 recruitment to SHPS-1. The role of Grb2 was confirmed using a Pyk2 Y881F mutant, which blocked Grb2 recruitment to SHPS-1. Cells expressing this mutant had impaired Nox4 recruitment to SHPS-1. IGF-I-stimulated downstream signaling and biological actions were also significantly impaired in Nox4 Y491F-overexpressing cells. Disruption of Nox4 recruitment to SHPS-1 in aorta from diabetic mice inhibited IGF-I-stimulated Src oxidation and activation as well as cell proliferation. These findings provide insight into the mechanism by which localized Nox4-derived ROS regulates the sustained activity of a tyrosine kinase that is critical for mediating signal transduction and biological actions.     

石刁柏胚乳愈伤组织的诱导及植株的再生

石刁柏已形成细胞的幼嫩胚乳,接种在附加有不同浓度的生长素(NAA)和细胞分裂素(BA)的 MS 培养基上,获得了愈伤组织。愈伤组织的诱导频率随生长素的浓度不同而异,可达65.9—83.1%。将胚乳愈伤组织转移到降低了生长素浓度或只含有低浓度生长素的分化培养基上,可陆续分化芽、根、芽丛和少量胚状体,个别的芽和胚状体能发育成小植株。切取1.5—5cm 长的芽,接种在诱导根的培养基上,或在 IBA50ppm 溶液中浸泡2小时,转移到 MS 基本培养基上,部分芽能生根形成完整植株。     

Gene mapping on maize pachytene chromosomes by in situ hybridization

A sensitive in situ hybridization technique which was effective for mapping genes of low copy number on human metaphase chromosomes was used for gene mapping on maize pachytene chromosomes. A cloned genomic EcoR1 fragment of 10.8 kb, containing most or all of the sequence encoding the Waxy locus mRNA, was used as the probe. Southern DNA blotting analyses performed by Shure et al. (1983) indicated that the Waxy locus was a single copy sequence. In our in situ hybridization experiment, the probe hybridized to a specific site on chromosome 9. Labeling at this site was detected in 48.6% of 154 randomly selected copies of chromosome 9. To test the sensitivity of the method, subclones of the fragment with insert sizes of 6.6, 4.7, 3.5, 2.3, 1.9 and 0.8 kb were used for in situ hybridizations. Labeling efficiency for each probe was determined. The data showed that a single copy probe of 1.9 kb could be detected at the correct position in 18% of 183 randomly selected number 9 chromosomes.     

苦绳的一个新甾体成分

从苦绳(Dregea sinensis Hemsl.),和分出一新的甾体成分,Dresigenin A(I),经化学反应及光谱分析证明其结构为12-O-乙酰基-20-O-(2-甲基丁酰基)二氢肉珊瑚甙元[12-O-acetyl-20-O-(2-methylbutyryl)dihydrosarcostin]。     

Substitution of an invariant nucleotide at the base of the highly conserved ''530-loop'' of 15S rRNA causes suppression of yeast mitochondrial ochre mutations.

We have determined the nucleotide sequence alteration in the 15S rRNA gene of a Saccharomyces cerevisiae strain carrying the previously described mitochondrial ochre suppressor, MSUI. The suppressor contains an A residue at position 633 of the yeast mitochondrial sequence, in place of the wild-type G. This position, located in the highly conserved region forming the stem of the '530-loop', corresponds to G517 of the Escherichia coli 16S rRNA and is occupied by G in all other known small rRNA sequences. This finding strongly supports the previous conclusions of others that the 530-loop region plays an important role in enhancing translational accuracy.     

A specific, photolabile and irreversible antagonist (L662,025) of the PAF-receptor

PAF (0.2 microM) induced maximal platelet aggregation in human PRP and [3H]-PAF (1-5 nM) binding to platelet membrane preparations had Kd value of 3.8 nM and Bmax of 200 fmoles/mg of protein. Without UV irradiation, a synthetic azido tetrahydrofuran derivative L662,025 was a reversible and competitive PAF-receptor antagonist with IC50 values of 5.6 +/- 0.3 microM (platelet aggregation) and 1.0 +/- 0.25 microM (receptor binding). Photolysis of L662,025 in the presence of PRP produced an irreversible inhibition of platelet aggregation and specific binding of [3H]-PAF (1 nM). L662,025 did not affect collagen- or ADP-induced human platelet aggregation before or after photolysis. It is a new probe that can be used to identify and characterize the PAF-receptor.     

香薷中的化学成分

从香[Eltholtzia ciliata(Thund.)Hyland]中分出14个化合物,用波谱和化学等方法确定为6-甲基三十三烷(Ⅰ),13-环己基二十六烷(Ⅱ),β-谷甾醇(Ⅲ),棕榈酸(Ⅳ_a),亚油酸(Ⅳ_b),亚麻酸(Ⅳ_c),熊果酸(Ⅴ),5-羟基-6、7-二甲氧基黄酮(Ⅵ),5-羟基-7、8-二甲氧基黄酮(Ⅶ),5、7-二羟基-4′-甲氧基黄酮(Ⅷ),5-羟基-7、4′-二甲氧基双氢黄酮醇(Ⅸ),β-谷甾醇-3-β-D-葡萄糖甙(Ⅹ),5-羟基-6-甲基-7-O-α-D-半乳吡喃糖双氢黄酮甙(Ⅺ),刺槐素-7-O-β-D-葡萄糖甙(Ⅻ)。其中化合物Ⅺ为新化合物,除(Ⅳ_b)和(Ⅳ_c)外,其余为首次从该属植物中得到。     

北京地区大葱和玉米根际VA菌根的季节变化及其与环境因子之间的关系

1986-1987年连续两年系统调查了北京地区大葱(Allium fistulosum L.)和玉米(Zeamays L.)根际 VA 菌根侵染百分率及菌根真菌孢子密度的季节变化。分析了菌根和菌根真菌的发生与几种土壤因子(土壤硝态氮、铵态氮、速效磷和有机质含量,土壤 pH 值)和气候因子(气温、积温、土温、光照、降水量,相对湿度等)之间的关系。结果表明:大葱和玉米的侵染百分率随作物生育期的增长而增加,并以6月和9月的增长速度最快。一年中,植物根际菌根真菌的孢子密度有二个高峰,分别在6-7月和10月。在北京的同一地区,一年中,几种测定的土壤因子变化很小,因此,认为气候因子对菌根发生的季节变化具有更大的影响。用线性相关和关联度分析的结果表明:一年中,菌根的发生与积温的关系最为密切。     

国产磨芋属的染色体核型报道(1)

本文报道了磨芋属(Amorphophallus Blume)六个种的染色体数目和核型,其中5个种属于首次报道。其核型公式如下: 1.滇磨芋 K(2n)=2x=26=26m.2.磨芋 K(2n)=2x=26=26m.3.攸落磨芋K(2n)=2x=26=22m(2SAT)+4sm.(2SAT).4.西盟磨芋 K(2n)=2x=26=20m+4sm+2st.5.勐海磨芋 K(2n)=2x=26=22m+4sm.6.白磨芋 K(2n)=2x=26=20m(2SAT)+6sm。