Qiuping Zhang

King's College London | KCL · Cardiovascular Division

Nesprins comprise a family of multi-isomeric scaffolding proteins, forming the linker of nucleoskeleton-and-cytoskeleton complex with lamin A/C, emerin and SUN1/2 at the nuclear envelope. Mutations in nesprin-1/-2 are associated with Emery-Dreifuss muscular dystrophy (EDMD) with conduction defects and dilated cardiomyopathy (DCM). We have previousl...

Nesprins (nuclear envelope spectrin repeat proteins) are multi-isomeric scaffolding proteins. Giant nesprin-1 and -2 localise to the outer nuclear membrane, interact with SUN (Sad1p/UNC-84) domain-containing proteins at the inner nuclear membrane to form the LInker of Nucleoskeleton and Cytoskeleton (LINC) complex, which, in association with lamin...

Introduction Cardiomyopathies are an important cause of heart failure and sudden cardiac death. Emerging evidence demonstrated the importance of the mechanical properties of cardiomyocytes as new causes for dilated cardiomyopathy (DCM). Nesprins-1/2 are highly expressed in cardiac and skeletal muscle and together with SUN1/2, lamin A/C and emerin f...

Vascular smooth muscle cells (VSMCs) are the predominant cell type in the blood vessel wall. Changes in VSMC actomyosin activity and morphology are prevalent in cardiovascular disease. The actin cytoskeleton actively defines cellular shape and the LInker of Nucleoskeleton and Cytoskeleton (LINC) complex, comprised of nesprin and the Sad1p, UNC-84 (...

Nesprins (nuclear envelope spectrin repeat proteins) are a family of multi-isomeric scaffolding proteins. Nesprins form the LInker of Nucleoskeleton-and-Cytoskeleton (LINC) complex with SUN (Sad1p/UNC84) domain-containing proteins at the nuclear envelope, in association with lamin A/C and emerin, linking the nucleoskeleton to the cytoskeleton. The...

Nesprins (nuclear envelope spectrin repeat proteins) are multi-isomeric scaffolding proteins. Nesprin-1 and -2 are highly expressed in skeletal and cardiac muscles and together with SUN (Sad1p/UNC84) domain-containing proteins form the LInker of Nucleoskeleton and Cytoskeleton (LINC) complex at the nuclear envelope in association with lamin A/C and...

Introduction Vascular calcification is a hallmark of vascular ageing, and associated with vascular smooth muscle cell (VSMC) death, phenotype modulation and maladaptation. However, it remains unclear how the initial stress signals link to these downstream cellular events. Emerging evidence and our in vitro data suggest that stress may drive vascula...

Nesprins-1 and -2 are highly expressed in skeletal and cardiac muscle and together with SUN (Sad1p/UNC84)-domain containing proteins and lamin A/C form the LInker of Nucleoskeleton-and-Cytoskeleton (LINC) bridging complex at the nuclear envelope (NE). Mutations in nesprin-1/2 have previously been found in patients with autosomal dominant Emery-Drei...

Aims: Although extracellular-regulated kinases (ERK) are a well-known central mediator in cardiac hypertrophy, no clinically available ERK antagonist has been tested for preventing cardiac hypertrophy. Selumetinib is a novel oral MEK inhibitor that is currently under Phase II and Phase III clinical investigation for advanced solid tumors. In this...

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Supplementary material

Background Nesprin-1-giant (1008kD) is a protein of the outer nuclear membrane that links nuclei to the actin cytoskeleton via amino-terminal calponin homology domains. The short nesprin-1 isoform, nesprin-1-α2, is present only in skeletal and cardiac muscle and several pathogenic mutations occur within it, but the functions of this short isoform w...

The spatial compartmentalisation of biochemical signalling pathways is essential for cell function. Nesprins are a multi-isomeric family of proteins that have emerged as signalling scaffolds, herein, we investigate the localisation and function of novel nesprin-2 N-terminal variants. We show that these nesprin-2 variants display cell specific distr...

Nesprins are a family of multi-isomeric scaffolding proteins that were originally identifi ed at the nuclear envelope (NE), where they bind to lamin A/C, emerin, and SUN-domain containing proteins, to form the LInker of Nucleoskeleton-and-Cytoskeleton (LINC) complex that connects the NE to the actin cytoskeleton. However, nesprin genes also give ri...

Prelamin A accumulation and persistent DNA damage response (DDR) are hallmarks of vascular smooth muscle cell (VSMC) ageing and dysfunction. Although prelamin A is proposed to interfere with DNA repair, our understanding of the crosstalk between prelamin A and the repair process remains limited. The extracellular signal-regulated kinases 1 and 2 (E...

Nesprins comprise a family of multi-isomeric scaffolding proteins that bind to lamin A/C, emerin and SUN1/2 at the nuclear envelope (NE) to form the linker of nucleoskeleton and cytoskeleton (LINC) complex. Mutations in nesprin-1 and -2 contribute to Emery–Dreifuss muscular dystrophy and dilated cardiomyopathy (DCM). Following mutation screening in...

Nesprin proteins have a central rod domain of spectrin repeats and are intracellular linkers and scaffolds. Full length or “giant” nesprin-1 and nesprin-2 have N-terminal calponin homology domains that bind the actin cytoskeleton and C-terminal transmembrane KASH domains which anchor the nesprins to the outer nuclear membrane. A number of short iso...

Aims: β-catenin has been shown to be regulated by inducible nitric oxide synthase (NOS) in endothelial cells. We investigated here whether β-catenin interacts with and regulates endothelial NOS (eNOS) and whether eNOS activation promotes β-catenin signalling. Methods and results: We identified β-catenin as a novel eNOS binding protein in human u...

Nesprin-1-giant and nesprin-2-giant regulate nuclear positioning by the interaction of their C-terminal KASH domains with nuclear membrane SUN proteins and their N-terminal calponin-homology domains with cytoskeletal actin. A number of short isoforms lacking the actin-binding domains are produced by internal promotion. We have evaluated the signifi...

Nitric oxide (NO) derived from endothelial NO synthase (eNOS) exerts cardioprotective effects. eNOS binds to a number of proteins that regulate its function. Using mass spectroscopy to study eNOS immunoprecipitated from human umbilical vein endothelial cells (HUVEC), we identified β-catenin as a novel binding partner of eNOS. This previously unreco...

A combination of isoform-specific, quantitative RT-PCR and site-specific monoclonal antibodies has been used to produce consistent evidence at both RNA and protein levels for the principal isoform products of SYNE1 and SYNE2, the genes encoding nesprin-1 and nesprin-2 proteins. Western blots show that both genes produce three groups of isoforms tha...

Nesprins (Nuclear envelope spectrin-repeat proteins) are a novel family of giant spectrin-repeat containing proteins. The nesprin-1 and nesprin-2 genes consist of 146 and 116 exons which encode proteins of ∼1mDa and ∼800 kDa is size respectively when all the exons are utilised in translation. However emerging data suggests that the nesprins have mu...

Localizations of p23Nesp1, p12Nesp1 and p41Nesp1. A) p23Nesp1, p12Nesp1 and p41Nesp1 displayed diffusive cytoplasmic localization when transfected into U2OS cells. B) p41Nesp1 displayed diffusive localization and concentrated around the ER when transfected into HDFs. (TIF)

Primers used for 5′ and 3′ RACE. Primers and nested primers used for detection of novel nesprin-1 and nesprin-2 cDNA ends. (DOCX)

Schematics of nesprin-1 and nesprin-2 cassette exons. A) Nesprin-1 genomic map showing the location of nesprin-1 cassette exon 6′ (Red box). B) Nesprin-2 genomic map showing the location of nesprin-2 cassette exons 101’ (orange box) and107’ (Purple box). (TIF)

UTR combinations used to generate potential nesprin-1 variants. Nesprin-1 can generate multiple variants through the use of alternative UTRs in a ‘mix-and-match’ approach. The tables highlight the UTR pairs used to generate the potential isoforms described in Figures 2B. (DOCX)

UTR combinations used to generate potential nesprin-2 variants. Nesprin-2 can generate multiple variants through the use of alternative UTRs in a ‘mix-and-match’ approach. The tables highlight the UTR pairs used to generate the potential isoforms described in Figures 3B. (DOCX)

Primers used for UTR detection. Forward and reverse primers used for detection of novel nesprin-1 and nesprin-2 UTRs. Forward and reverse primers were separated by at least 1 coding exon to control for genomic contamination. (DOCX)

Primers used for the detection of ΔKASH variants. Forward and reverse primers used for detection of nesprin-1 and nesprin-2 ΔKASH variants. (DOCX)

Primers used for detection of cassette exons. Forward and reverse primers used for detection of novel nesprin-1 and nesprin-2 cassette exons. (DOCX)

Lamin A/C mutations are the most common cause of familial dilated cardiomyopathy (DCM) but the pathogenetic mechanisms are incompletely understood. Nesprins are spectrin repeat-containing proteins that interact with lamin A/C and are components of the linker-of-nucleoskeleton-and-cytoskeleton (LINC) complex that connects the nuclear envelope to the...

Objective To investigate the role of Nesprin-1, a novel spectrin repeat containing scaffolding protein, in skeletal muscle cell differentiation. Methods During in vitro differentiation of C2C12 myoblasts into myotubes, changes in Nesprin-l gene expression and sub-cellular localization were detected using Western blotting and immunofluorescence. RNA...

Mendelian susceptibility to mycobacterial disease (MSMD) is a rare primary immunodeficiency associated with clinical disease caused by weakly virulent mycobacterial species. Interferon gamma receptor 1 (IFN-gammaR1) deficiency is a genetic etiology of MSMD. We describe the clinical and genetic features of a 7-year-old Italian boy suffering from MSM...

Nuclear and cytoplasmic scaffold proteins have been shown to be essential for temporal and spatial organization, as well as the fidelity, of MAPK signaling pathways. In this study we show that nesprin-2 is a novel extracellular signal-regulated MAPK1 and 2 (ERK1/2) scaffold protein that serves to regulate nuclear signaling by tethering these kinase...

Background: Epidermal growth factor receptor (EGFR) has been recently implicated in pathological tissue remodelling and sustained remodelling processes can lead to pathological outcomes, such as cardiac hypertrophy in heart failure. Dilated cardiomyopathy (DCM) is the most common form of heart muscle disease, comprising 60% of the cases of identif...

Emery–Dreifuss muscular dystrophy (EDMD) is a heterogeneous late-onset disease involving skeletal muscle wasting and heart defects caused, in a minority of cases, by mutations in either of two genes encoding the inner nuclear membrane (INM) proteins, emerin and lamins A/C. Nesprin-1 and -2 are multi-isomeric, spectrin-repeat proteins that bind both...

Emerin and specific isoforms of nesprin-1 and -2 are nuclear membrane proteins which are binding partners in multi-protein complexes spanning the nuclear envelope. We report here the characterisation of the residues both in emerin and in nesprin-1alpha and -2beta which are involved in their interaction and show that emerin requires nesprin-1 or -2...

Mutations of the nuclear lamins cause a wide range of human diseases, including Emery-Dreifuss muscular dystrophy and Hutchinson-Gilford progeria syndrome. Defects in A-type lamins reduce nuclear structural integrity and affect transcriptional regulation, but few data exist on the biological role of B-type lamins. To assess the functional importanc...

Nesprins are a recently discovered family of ubiquitously expressed intracellular proteins. Through alternative transcriptional initiation, termination and splicing, two genes - nesprin-1 and nesprin-2 (also known as syne-1 and syne-2) - give rise to many protein isoforms that vary markedly in size. The largest of these isoforms comprise a C-termin...

Nesprin-2 is a multi-isomeric, modular protein composed of variable numbers of spectrin-repeats linked to a C-terminal transmembrane domain and/or to N-terminal paired calponin homology (CH) domains. The smaller isoforms of nesprin-2 co-localize with and bind lamin A and emerin at the inner nuclear envelope (NE). In SW-13 cells, which lack lamin A/...

Nesprin-2 is a multi-isomeric, modular protein composed of variable numbers of spectrin-repeats linked to a C-terminal transmembrane domain and/or to N-terminal paired calponin homology (CH) domains. The smaller isoforms of nesprin-2 co-localize with and bind lamin A and emerin at the inner nuclear envelope (NE). In SW-13 cells, which lack lamin A/...

Mineralization-regulating proteins are found deposited at sites of vascular calcification. However, the relationship between the onset of calcification in vivo and the expression of genes encoding mineralization-regulating proteins is unknown. This study aimed to determine the temporal and spatial pattern of expression of key bone and cartilage pro...

Nesprin-1 and nesprin-2 (also known as Syne-1 and Syne-2,) are large ( approximately 3300-residue) vertebrate proteins associated with emerin and lamin A at the nuclear envelope of muscle cells and other cell types. We show that the previously described nesprins are short isoforms of giant proteins comprising an actin-binding amino-terminus connect...

Fifteen polymorphisms in six lipid transport genes were studied in a German population for relationships with dyslipidemia and coronary artery disease (CAD), to investigate a possible genetic basis for the marked differences in mortality rates from coronary heart disease within Europe. In other populations these polymorphisms have all been associat...

In search of vascular smooth muscle cell differentiation markers, we identified two genes encoding members of a new family of type II integral membrane proteins. Both are ubiquitously expressed, and tissue-specific alternative mRNA initiation and splicing generate at least two major isoforms of each protein, with the smaller isoforms being truncate...

The large ethnic differences in prevalence of coronary artery disease between China and Europe may relate to both genetic and environmental differences. To assess possible genetic factors we have therefore studied the frequencies of disease-related variants of genes involved in lipid transport in 69 hypertriglyceridemic Chinese subjects and 74 heal...

1. The common association of obesity, diabetes mellitus and hyperlipidaemia may have a primary aetiological basis. Insulin resistance has been postulated as a possible cause, but defects in the plasma transport of triacylglycerol or fatty acids could also be primary determinants. 2. We have therefore, studied 18 patients with diabetes mellitus, obe...

The restriction fragment length polymorphisms (RFLPs) of apolipoprotein(apo) A I gene were studied using polymerase chain reaction (PCR) in 69 endogenous hypertriglyceridemics (HTG) and 74 healthy subjects from a population of Chinese Han nationality in Chengdu area. The loci studied included Msp 1 within intron 3 of the apo A I gene; Xmn I, 5' to...

Sequencing analysis of lipoprotein lipase (LPL) gene exons l-9 was made in 11 cases of Chinese patients with severe endogenous hypertriglyceridemia (serum TG > 7.68 mmol/L) by the dideoxy chain termination method using Sequenase PCR Product Sequencing Kit. Four of the patients were found to possess 3 variants at the LPL gene locus. One novel mutati...

Two common coding sequence mutations of lipoprotein lipase (serine447-ter, producing a carboxy terminal truncation; and asp9-asn variants) were studied in 329 Caucasian subjects, of whom 243 had angiographic features of premature atheroscelerosis (220 with coronary artery disease; 23 with coronary and peripheral artery disease). As expected, the me...

A Finnish kindred with premature coronary heart disease and decreased HDL cholesterol levels was identified as having an apoA-I variant, apoA-I (Lys107-->0), caused by a 3-bp deletion of nucleotides 1396 through 1398 in exon 4 of the apoA-I gene. These subjects (n = 10) were heterozygous for this mutation. The mean serum HDL cholesterol concentrati...