Scaffolds,levers, rods and springs: diverse cellular functions of long coiled-coil proteins

Long alpha-helical coiled-coil proteins are involved in a variety of organizational and regulatory processes in eukaryotic cells. They provide cables and networks in the cyto- and nucleoskeleton, molecular scaffolds that organize membrane systems, motors, levers, rotating arms and possibly springs. A growing number of human diseases are found to be caused by mutations in long coiled-coil proteins. This review summarizes our current understanding of the multifaceted group of long coiled-coil proteins in the cytoskeleton, nucleus, Golgi and cell division apparatus. The biophysical features of coiled-coil domains provide first clues toward their contribution to the diverse protein functions and promise potential future applications in the area of nanotechnology. Combining the power of fully sequenced genomes and structure prediction algorithms, it is now possible to comprehensively summarize and compare the complete inventory of coiled-coil proteins of different organisms.Received 27 January 2004; received after revision 23 February 2004; accepted 10 March 2004     

Menadione-induced apoptosis and its mechanism in plants

Menadione can induce apoptosis in tobacco protoplasts. Typical characteristics are detected including the condensation of chromatin, the formation of apoptotic bodies and the degradation of genomic DNA into “DNA ladder”. Specific DNase is activated during this process. Ca²⁺ and Mg²⁺ are necessary for its activation, while Zn²⁺ and EDTA (Ethylenediaminetetraacetic acid) can inhibit its activation. The fragmentation of DNA and lamin can be inhibited by DEVD (Ac-Asp -Glu- Val- Asp-aldehyde). The fragmentation of lamin can also be inhibited by PMSF (Phenylmethylsulfonyl fluoride) and CH (Cyclohexinide). These results show that activation of specific DNase and proteases is involved in menadion-induced apoptosis in plants.     

Menadione-induced apoptosis and the degradation of lamin-like proteins in tobacco protoplasts

Detection of stereotypic hallmarks of apoptosis during cell death induced by menadione, including DNA laddering and the formation of apoptotic bodies, is reported. Comet assay and the TdT-mediated dUTP nick end labelling (TUNEL) procedure were also performed to detect DNA fragmentation. Inhibition of DNA fragmentation by Ac-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO) and phenylmethylsulfosyl (PMSF) implicated the involvement of caspase-like proteases in menadione-induced apoptosis in plants. We further studied the cleavage of lamin-like proteins during apoptosis in menadione-treated tobacco protoplasts. In animals, it has been reported that the solubilization of nuclear lamina and lamin degradation occurs during apoptotic cell death. However, little is known about the fate of lamins in apoptotic plant cells. Our study provided evidence that lamin-like proteins degraded into 35-kDa fragments in tobacco protoplasts induced by menadione, and this preceded DNA fragmentation. The results thus indicated that proteolytic cleavage of nuclear lamins was also conserved in programmed cell death in plants. Received 16 November 1998; received after revision 21 December 1998; accepted 23 December 1998     

Lamins and their genes

The nuclear lamina is an important nuclear structure. The studies on its peptides lamins and genes have made substantial progress in recent years. In this paper, the new achievements in studies of new lamin members and their functions, lamin genes and their origin and differentiation, are reviewed and discussed. Some research areas that deserve to be investigated further are put forward.     

Emery-Dreifuss muscular dystrophy at the nuclear envelope: 10 years on

Emery-Dreifuss muscular dystrophy (EDMD) is a neuromuscular degenerative condition with an associated dilated cardiomyopathy and cardiac conduction defect. It can be inherited in either an X-linked or autosomal manner by mutations in the nuclear proteins emerin and lamin A/C, respectively. Traditionally muscular dystrophies were associated with defects in sarcolemma-associated proteins and, therefore, a nuclear connection suggested the existence of novel signalling pathways associated with this group of diseases. Subsequently, other mutations in the lamin A/C gene were attributed to a range of tissue-specific degenerative conditions, collectively known as the ‘laminopathies’. Therefore, any proposed hypothesis underlying the molecular mechanism of EDMD needs to include this anomaly. As we celebrate the 10th anniversary of the identification of emerin as a component of the nuclear envelope, I discuss here the available evidence that currently implicates EDMD as arising from perturbations in myogenic regulatory pathways, causing temporal delays in both cell cycle progression and muscle regeneration. Received 25 May 2006; received after revision 22 June 2006; accepted 22 August 2006     

Human progeroid syndromes,aging and cancer: new genetic and epigenetic insights into old questions

Disorders in which individuals exhibit certain features of aging early in life are referred to as segmental progeroid syndromes. With the progress that has been made in understanding the etiologies of these conditions in the past decade, potential therapeutic options have begun to move from the realm of improbability to initial stages of testing. Among these syndromes, relevant advances have recently been made in Werner syndrome, one of several progeroid syndromes characterized by defective DNA helicases, and Hutchinson-Gilford progeria syndrome, which is characterized by aberrant processing of the nuclear envelope protein lamin A. Although best known for their causative roles in these illnesses, Werner protein and lamin A have also recently emerged as key players vulnerable to epigenetic changes that contribute to tumorigenesis and aging. These advances further demonstrate that understanding progeroid syndromes and introducing adequate treatments will not only prove beneficial to patients suffering from these dramatic diseases, but will also provide new mechanistic insights into cancer and normal aging processes. Received 28 July 2006; received after revision 5 September 2006; accepted 13 October 2006     

几类低等生物的核纤层与核纤层的起源进化研究

综合分析了本实验室多年来在几类具有重要进化地位的低等单位真核生物上进行的核纤层结构与成分的研究结果,并结合发育生物学等方面的资料,认为核纤层结构在真核细胞起源进化的初期即已起源形成,它的起源形成是“原核”进化成“真核”的重要前提条件之一;核纤层蛋白的起源分化过程应该是：首先起源产生Ｂ型蛋白,在此基础上分化出Ａ型蛋白,最后形成了现存高等真核细胞的核纤层蛋白家族。     

螅状独缩虫(Carchesium polypinum)类中间纤维的初步研究

利用间接免疫荧光和免疫印迹发现螅状独缩虫中存在两种分子质量为58ku和66ku的蛋白。这两种蛋白能分别与抗波形蛋白和抗核纤层蛋白B的抗体反应。分子质量66ku的蛋白位于大核边缘。另外,细胞经分级抽提后大核周围存在核纤层样结构。基于上述结果,可以认为螅状独缩虫中存在类中间纤维。     

草履虫类核纤层蛋白的初步研究

本文以DGD包埋去包埋技术对草履虫的核纤层通过透射电镜和免疫荧光、分子杂交等技术进行了观察。结果显示,在核周存在由10nm纤维组成的核纤层;免疫荧光结果表明,在核周呈阳性反应,并在其表皮、口器等部位呈交叉的阳性反应;蛋白分子杂交的反应带在68kD处呈阳性反应。