The nuclear pore complex (NPC) and its relationship to the envelope (NE) was characterized in living cells using POM121-green fluorescent protein (GFP) GFP-Nup153, GFP-lamin B1. No independent movement of single complexes found within plane NE interphase. Only large arrays NPCs moved slowly synchronously during global changes shape, strongly suggesting mechanical connections which form an NPC network. lamina exhibited identical movements. turnover measured by fluorescence recovery after...

We have identified an integral membrane protein of 145 kD (estimated by SDS-PAGE) rat liver nuclear envelopes that binds to WGA. obtained peptide sequence from purified p145 and cloned sequenced several cDNA clones one genomic clone. The relative molecular mass calculated its complete, deduced primary structure is 120.7 kD. Antibodies raised against a synthetic represented in reacted monospecifically with p145. In indirect immunofluorescence these antibodies gave punctate staining the...

ABSTRACT Emerin is an integral protein of the inner nuclear membrane that mutated or not expressed in patients with Emery-Dreifuss muscular dystrophy. Confocal immunofluorescence microscopy studies intracellular targeting truncated forms emerin, some which are found dystrophy, show nucleoplasmic, amino-terminal domain necessary and sufficient for retention. When this fused to a transmembrane segment ER/plasma membrane, chimeric localized membrane. The emerin targeted Fluorescence...

Proteins with molecular masses ranging from 30 kDa. (green fluorescent protein, GFP) to 150 kDa (monoclonal and polyclonal antibodies) were coupled the cellular translocating peptide transportan ...

The HIV-1 genome contains several genes coding for auxiliary proteins, including the small Vpr protein. affects integrity of nuclear envelope and participates in translocation preintegration complex containing viral DNA. Here, we show by photobleaching experiments performed on living cells expressing a Vpr-green fluorescent protein fusion that shuttles between nucleus cytoplasm, but significant fraction is concentrated at envelope, supporting hypothesis interacts with components pore...

We have produced new antibodies specific for the integral pore membrane protein POM121. Using these we show that during apoptosis POM121 becomes proteolytically degraded in a caspase-dependent manner. The and other proteins of nuclear envelope were used comparative study staurosporine-treated buffalo rat liver cells. Nuclei from cells classified three different stages apoptotic progression: stage I, moderately condensed chromatin surrounded by smooth periphery; II, compact patches collapsing...

Here, we characterize a transmembrane protein of the nuclear envelope that name spindle-associated membrane 1 (Samp1). The is conserved in metazoa and fission yeast homologous to Net5 rat Ima1 Schizosaccharomyces pombe. We show that, human cells, membrane-spanning polypeptide with an apparent molecular mass 43 kDa. This consistent predicted 392 amino acids has five segments its C-terminus exposed nucleoplasm. During interphase, Samp1 was specifically distributed inner membrane....

c-Myc is a predominately nuclear transcription factor that substrate for rapid turnover by the proteasome system. Cancer-related mutations in lead to defects its degradation and thereby contribute increase cellular level associated with disease. Little known about mechanisms target proteasomes. By using GFP fusion protein live analysis we show shuttles between nucleus cytoplasm thus it could be degraded either compartment. Strikingly, at elevated levels of expression accumulates nucleoli...

The transmembrane inner nuclear membrane (INM) protein Samp1 is required for anchoring centrosomes near the nuclei. Using high-resolution fluorescence microscopy we show that distributed in a distinct and characteristic pattern envelope (NE), where it partially colocalizes with LINC complex Sun1. By studying localization of deletion mutants fusion proteins, conclude cysteine-rich N-terminal half nucleoplasmically exposed responsible targeting to INM. It contains four conserved CxxC motifs...

Disassembly and reassembly of the nuclear pore complexes (NPCs) is one major events during open mitosis in higher eukaryotes. However, how this process controlled by mitotic machinery not clear. To investigate we developed a novel vivo model system based on syncytial Drosophila embryos. We microinjected different effectors into embryonic cytoplasm monitored dynamics disassembly/reassembly NPCs live embryos using fluorescently labeled wheat germ agglutinin (WGA) or fixed electron microscopy...

Cell-penetrating peptides (CPPs) uptake mechanism is still in need of more clarification to have a better understanding their action the mediation oligonucleotide transfection. In this study, effect on early events (1 h treatment) transfection by PepFect14 (PF14), with or without cargo gene expression, HeLa cells, been investigated. The RNA expression profile was characterized sequencing and confirmed qPCR analysis. regulations were then related biological processes study signaling pathways...

ABSTRACT We have investigated a possible role for the inner nuclear membrane protein Samp1 (also known as TMEM201) in mitotic machinery. Live-cell imaging showed that Samp1a–YFP (Samp1a is short isoform of Samp1) distributed filamentous structures spindle, partially colocalising with β-tubulin. depletion resulted an increased frequency cells signs chromosomal mis-segregation and prolonged metaphase, indicating problems spindle assembly and/or alignment. Consistent this, spindles...

Investigating interactions of proteins in the nuclear envelope (NE) using co-immunoprecipitation (Co-IP) has previously been difficult or even impossible due to their inherent resistance extraction. We have developed a novel method, MCLIP (Membrane protein Cross-Link ImmunoPrecipitation), which takes advantage cell permeable crosslinker enable effective detection and analysis specific NE live cells Western blot. Using we show that, U2OS cells, integral inner membrane Samp1 interacts with...

Polyglutamine (polyQ) diseases, such as Spinocerebellar ataxia type 7 (SCA7), are caused by expansions of polyQ repeats in disease specific proteins. The sequestration vital proteins into aggregates formed is believed to be a common pathological mechanism these disorders. RNA-binding protein FUS has been observed aggregates, though if disruption this plays role the neuronal dysfunction SCA7 or other diseases remains unclear. We therefore analysed localisation and function stable inducible...

The 121–kDa pore membrane protein (POM121) is a bitopic integral specifically located in the domain of nuclear envelope with its short N‐terminal tail exposed on luminal side and major C‐terminal portion adjoining complex. In order to locate signal for targeting POM121 pores, we over expressed selected regions alone or fused green fluorescent (GFP) transiently transfected COS‐1 cells stably neuroblastoma cell line. Microscopic analysis GFP fluorescence immunostaining was used determine...

The nuclear pore complexes (NPCs), multiprotein assemblies embedded in the envelope, conduct nucleo‐cytoplasmic traffic of macromolecules. Mimics NPCs, called annulate lamellae (ALPCs), are usually found cytoplasmic membranous stacks oocytes and early embryonic cells. They believed to constitute storage compartments for excess premade nucleoporins. To evaluate extent which ALPCs store nucleoporins cells we took advantage syncytial Drosophila embryos, containing both AL rapidly proliferating...

Gp210, an integral membrane protein of the nuclear pore complex (NPC), is believed to be involved in NPC biogenesis. To test this hypothesis, we have investigated dynamic properties and distribution proteins NIH/3T3 cells lacking gp210. POM121 (the other protein) NUP107 (of NUP107/160 complex) were correctly distributed at pores absence Furthermore, fluorescence recovery after photobleaching experiments showed that remained stably associated NPCs. We conclude gp210 cannot required for...