Expansion of the ISWI chromatin remodeler family with new active complexes

ISWI chromatin remodelers mobilize nucleosomes to control DNA accessibility. Complexes isolated to date pair one of six regulatory subunits with one of two highly similar ATPases. However, we find that each endogenously expressed ATPase co‐purifies with every regulatory subunit, substantially increasing the diversity of ISWI complexes, and we additionally identify BAZ2B as a novel, seventh regulatory subunit. Through reconstitution of catalytically active human ISWI complexes, we demonstrate that the new interactions described here are stable and direct. Finally, we profile the nucleosome remodeling functions of the now expanded family of ISWI chromatin remodelers. By revealing the combinatorial nature of ISWI complexes, we provide a basis for better understanding ISWI function in normal settings and disease.     

Reduced expression of PMCA1 is associated with increased blood pressure with age which is preceded by remodelling of resistance arteries

Hypertension is a well‐established risk factor for adverse cardiovascular events, and older age is a risk factor for the development of hypertension. Genomewide association studies have linked ATP2B1, the gene for the plasma membrane calcium ATPase 1 (PMCA1), to blood pressure (BP) and hypertension. Here, we present the effects of reduction in the expression of PMCA1 on BP and small artery structure and function when combined with advancing age. Heterozygous PMCA1 null mice (PMCA1^Ht) were generated and conscious BP was measured at 6 to 18 months of age. Passive and active properties of isolated small mesenteric arteries were examined by pressure myography. PMCA1^Ht mice exhibited normal BP at 6 and 9 months of age but developed significantly elevated BP when compared to age‐matched wild‐type controls at ≥12 months of age. Decreased lumen diameter, increased wall thickness and increased wall:lumen ratio were observed in small mesenteric arteries from animals 9 months of age and older, indicative of eutrophic remodelling. Increases in mesenteric artery intrinsic tone and global intracellular calcium were evident in animals at both 6 and 18 months of age. Thus, decreased expression of PMCA1 is associated with increased BP when combined with advancing age. Changes in arterial structure precede the elevation of BP. Pathways involving PMCA1 may be a novel target for BP regulation in the elderly.     

荞麦柱头、花柱的结构及ATP酶的超微细胞化学定位

利用超薄切片结合ATP酶定位技术,研究了荞科柱头及花柱的结构。结果表明,成熟的荞麦柱头表面无乳突细胞,柱头细胞的细胞壁存在发达的壁内突,花柱为实心型无引导组织,柱头表面的细胞壁根据电子密度的不同可划分为3层,最外一层电子密度最大,最内一层电子近透明,并形成壁内突的结构,柱头表面和花柱细胞的质膜上ATP酶的活性较高,表明成熟的柱头和花柱细胞物质代谢和运输十分活跃,成熟的柱头和花柱细胞中有丰富的蛋白质和多糖类物质。     

水稻液泡ATPaseB亚基基因的克隆及其在低磷胁迫下的表达特征分析(英)

利用抑制性扣除杂交 (SSH)技术构建水稻 (OryzasativaL .)根系磷饥饿诱导cDNA文库 ,获得编码液泡ATPase (V_ATPase)B亚基的克隆 ,通过反转录PCR方法获得该基因的完整序列。该基因编码 4 87个氨基酸 ,含有一个保守的ATP结合位点 ,其蛋白分子量为 5 4 .0 6kD ,等电点为 4 .99。Southern印迹表明 ,V_ATPaseB亚基基因在水稻基因组中以单拷贝形式存在。氨基酸同源性分析发现 ,V_ATPaseB亚基是一个较为保守的蛋白亚基 ,其序列变化伴随生物的进化过程同步进行。Northern印迹表明 ,V_ATPaseB亚基在水稻根系中受到磷饥饿诱导表达 ,磷饥饿 6～ 12h出现表达高峰 ,而在叶片中表达高峰有所滞后 (2 4～ 4 8h)。在缺磷环境条件下 ,ATPaseB亚基可能通过提高其表达量 ,进而提高质子转运活性 ,形成跨膜的电化学梯度 ,为体内储备磷跨液泡膜运输提供能量 ,从而提高植物体内磷的利用效率及其耐低磷的能力。     

毛竹茎秆纤维细胞发育过程中ATP酶的超微细胞化学定位研究

采用磷酸铅沉淀技术,对毛竹茎秆纤维细胞发育过程中的ATP酶进行了超微细胞化学定位研究.在初生壁形成时期,大量的ATP酶的活性产物沉积在质膜、质膜内陷、运输小泡、胞间连丝等膜体系以及细胞核和各种细胞器上;在次生壁形成的初期,ATP酶在多泡小体和裂解的液泡膜上出现,凝聚并边缘化的染色质上仍然具有ATP酶活性;随着次生壁的逐渐加厚,在前四年中持续存在具有ATP酶活性的质膜内陷结构,以后消失;而在六年生纤维细胞的质膜、运输小泡、纹孔、胞间连丝和凝聚化的染色质上仍然发现有明显的ATP酶分布,并发现在染色质上ATP酶活性会随着凝聚程度的加深而增强.结果表明,ATP酶在毛竹茎秆纤维细胞壁的整个形成过程中发挥重要作用,而纤维细胞的次生壁形成过程是一个由核基因控制的主动的PCD过程;并证实毛竹茎秆纤维细胞的发育有别于其它木本植物纤维细胞的发育过程,这种纤维细胞是一种典型的长寿细胞.     

冷驯化提高金边卫矛抗寒力期间细胞ATP酶的超微细胞化学定位

采用磷酸铅沉淀的电镜细胞化学方法,对经冷驯化及未经冷驯化的金边卫矛 EuonymusradicansEmorald&Gold. 以及两者在-5℃下处理1d后的叶肉细胞进行了ATPase的超微细胞化学定位的比较观察,并对冷驯化可提高植物抗寒力及ATPase与植物抗寒性关系进行了探讨.标志ATPase活性反应的磷酸铅沉淀物主要分布在叶片栅栏细胞和海绵细胞的质膜和液泡膜上,ATPase在这两类细胞中的分布及活性无明显差异.金边卫矛幼苗经4℃低温驯化7d后,其质膜和液泡膜的ATPase活性高于22℃下生长的幼苗,冷驯化后的金边卫矛幼苗在-5℃处理1d后,其质膜和液泡膜上仍可观察到ATPase的活性反应,但较低温胁迫前有所降低,且超微结构完整,未见任何伤害.而未经冷驯化的金边卫矛幼苗在-5℃处理1d后,其质膜和液泡膜上的ATPase完全失活,同时细胞的超微结构受到严重伤害.实验结果表明,冷驯化可提高金边卫矛幼苗ATPase在低温胁迫下的稳定性,这种活性变化与植物抗寒力的提高呈正相关,同时还发现液泡膜ATPase活性较质膜ATPase变化幅度大,推测液泡膜可能较质膜ATPase对低温更为敏感.     

The role of calcium channels in anthocyanin production in callus cultures of Daucus carota

The involvement of Ca²⁺ ATPases in anthocyanin accumulation in callus cultures of Daucus carota was investigated under the influence of calcium and calcium channel modulators. Ionophore (I) treatment enhanced callus growth and anthocyanin accumulation. Increasing the amount of calcium applied to cultures enhanced the anthocyanin level. Ionophore treatment influenced the enhancement of Ca²⁺ATPase and endogenous titres of PAs. Addition of the calcium channel blocker verapamil or the calmodulin antagonist chlorpromazine to the A23187 (ionophore) treated cells caused a reduction in anthocyanin levels. Channel blockers reduced Ca²⁺ATPase activity, which was restored by ionophore treatment, showing the importance of calcium in anthocyanin production. Higher ethylene levels were also found in treatment with ionophore or 2X calcium. Thus the influence of ionophore in anthocyanin production and its inhibition by calcium channel modulators suggests that calcium plays an important role in the production of anthocyanin by carrot callus cultures.