Nuclear Machinery Research Articles

FLN-2 functions in parallel to linker of nucleoskeleton and cytoskeleton complexes and CDC-42/actin pathways during P-cell nuclear migration through constricted spaces in Caenorhabditis elegans.

Nuclear migration through narrow constrictions is important for development, metastasis, and proinflammatory responses. Studies performed in tissue culture cells have implicated linker of nucleoskeleton and cytoskeleton (LINC) complexes, microtubule motors, the actin cytoskeleton, and nuclear envelope repair machinery as important mediators of nuclear movements through constricted spaces. However, little is understood about how these mechanisms operate to move nuclei in vivo. In Caenorhabditis elegans larvae, six pairs of hypodermal P cells migrate from lateral to ventral positions through a constricted space between the body wall muscles and the cuticle. P-cell nuclear migration is mediated in part by LINC complexes using a microtubule-based pathway and by an independent CDC-42/actin-based pathway. However, when both LINC complex and actin-based pathways are knocked out, many nuclei still migrate, suggesting the existence of additional pathways. Here, we show that FLN-2 functions in a third pathway to mediate P-cell nuclear migration. The predicted N-terminal actin-binding domain in FLN-2 that is found in canonical filamins is dispensable for FLN-2 function; this and structural predictions suggest that FLN-2 does not function as a filamin. The immunoglobulin-like repeats 4-8 of FLN-2 were necessary for P-cell nuclear migration. Furthermore, in the absence of the LINC complex component unc-84, fln-2 mutants had an increase in P-cell nuclear rupture. We conclude that FLN-2 functions to maintain the integrity of the nuclear envelope in parallel with the LINC complex and CDC-42/actin-based pathways to move P-cell nuclei through constricted spaces.

Evaluation of salinity-mediated end-point cytogenotoxicity in Germinating Roots of Lathyrus sativus L., Variety Mahatora.: Bio-assay guided biomarker studies

Pulse crops are susceptible to salt stress as per different research reports but how far Lathyrus sativus L., responds to increasing salinity has been taken up in this work. Thus, the harmful effects of increasing salinity on plant cells at various phases of chromosomal integrity and nucleolus morphology have been evaluated in Lathyrus sativus L., variety Mahatora. Lathyrus sativus variety Mahatora seeds were subjected to seed priming with serially diluted concentrations of NaCl (500, 400, 300, 200 and 100 mM respectively) and germination percentage (72 hrs), root length inhibition (7 days) normal and abnormal MI (Mitotic Index) with 2% aceto-orcein staining, nucleolar morphometric cum frequency analysis (0.05% hematoxylin), total protein, POX and Electrolyte leakage from etiolated roots and root metabolic activity/dehydrogenase activity were measured (TTC staining). From 200 mM onwards, significant reduction in germination percentage and root length inhibition resulted and at 300 and 400 mM salt-priming significant reduction in normal MI%, increased Abnormal MI% showing both aneugenic and clastogenic responses were accounted. At 500 mM pre-exposed root tip cells were found to develop gradual blackening and root tip death and very less viable cells with highly necrotic, vacuolated with chromosomal erosions and nuclear dismantling and nuclear blobbing resulted apoptosis in addition to decreased POX and dehydrogenase activity (300–500 mM NaCl-treated test sets). NaCl stands out as a potential cyto-genotoxicant in Lathyrus sativus L., variety Mahatora. The maximum tolerance level (200–300 mM) and at 400–500 mM NaCl has been highly cytotoxic as per cytological and biochemical data. From 200 mM onwards, nucleolar volume and frequency were altered and at 500 mM pretreatment complete degradation of nuclear machinery was encountered. Out of salinity significant proportions of C-mitosis and polyploidy showing NaCl has a disruptive action in spindle fibre formation that in turn produced somatic diads and subsequent polyploidy formations (At 200 to 300 mM).

Redox regulation of gene expression: proteomics reveals multiple previously undescribed redox-sensitive cysteines in transcription complexes and chromatin modifiers.

Redox signalling is crucial for regulating plant development and adaptation to environmental changes. Proteins with redox-sensitive cysteines can sense oxidative stress and modulate their functions. Recent proteomics efforts have comprehensively mapped the proteins targeted by oxidative modifications. The nucleus, the epicentre of transcriptional reprogramming, contains a large number of proteins that control gene expression. Specific redox-sensitive transcription factors have long been recognized as key players in decoding redox signals in the nucleus and thus in regulating transcriptional responses. Consequently, the redox regulation of the nuclear transcription machinery and its cofactors has received less attention. In this review, we screened proteomic datasets for redox-sensitive cysteines on proteins of the core transcription complexes and chromatin modifiers in Arabidopsis thaliana. Our analysis indicates that redox regulation affects every step of gene transcription, from initiation to elongation and termination. We report previously undescribed redox-sensitive subunits in transcription complexes and discuss the emerging challenges in unravelling the landscape of redox-regulated processes involved in nuclear gene transcription.

Mitochondrial stress activates YAP/TAZ through RhoA oxidation to promote liver injury

Yes-associated protein (YAP) and WW domain-containing transcription regulator protein 1 (WWTR1; also known as TAZ) are the main effectors of the Hippo pathway and their dysregulation contributes to diseases in tissues including the liver. Although mitochondria are capable of transmitting signals to change transcriptomic landscape of diseased hepatocytes, such retrograde signaling and the related nuclear machinery are largely unknown. Here, we show that increased YAP activity is associated with mitochondrial stress during liver injury; and this is required for secondary inflammation, promoting hepatocyte death. Mitochondrial stress inducers robustly promoted YAP/TAZ dephosphorylation, nuclear accumulation, and target gene transcription. RNA sequencing revealed that the majority of mitochondrial stress transcripts required YAP/TAZ. Mechanistically, direct oxidation of RhoA by mitochondrial superoxide was responsible for PP2A-mediated YAP/TAZ dephosphorylation providing a novel physiological input for the Hippo pathway. Hepatocyte-specific Yap/Taz ablation suppressed acetaminophen-induced liver injury and blunted transcriptomic changes associated with the pathology. Our observations uncover unappreciated pathway of mitochondrial stress signaling and reveal YAP/TAZ activation as the mechanistic basis for liver injury progression.

Nuclear Fusion in Yeast and Plant Reproduction.

Nuclear fusion is essential for the sexual reproduction of various organisms, including plants, animals, and fungi. During the life cycle of flowering plants, nuclear fusion occurs three times: once during female gametogenesis and twice during double fertilization, when two sperm cells fertilize the egg and the central cell. Haploid nuclei migrate in an actin filament-dependent manner to become in close contact and, then, two nuclei fuse. The nuclear fusion process in plant reproduction is achieved through sequential nuclear membrane fusion events. Recent molecular genetic analyses using Arabidopsis thaliana showed the conservation of nuclear membrane fusion machinery between plants and the budding yeast Saccharomyces cerevisiae. These include the heat-shock protein 70 in the endoplasmic reticulum and the conserved nuclear membrane proteins. Analyses of the A. thaliana mutants of these components show that the completion of the sperm nuclear fusion at fertilization is essential for proper embryo and endosperm development.

Understanding G-Quadruplex Biology and Stability Using Single-Molecule Techniques.

The link between the chemical stability of G-quadruplex (qDNA) structures and their roles in eukaryotic genomic maintenance processes has been an area of interest now for several decades. This Review seeks to demonstrate how single-molecule force-based techniques can provide insight into the mechanical stabilities of a variety of qDNA structures as well as their ability to interconvert between different conformations under conditions of stress. Atomic force microscopy (AFM) and magnetic and optical tweezers have been the primary tools used in these investigations and have been used to examine both free and ligand-stabilized G-quadruplex structures. These studies have shown that the degree of stabilization of G-quadruplex structures has a significant effect on the ability of nuclear machinery to bypass these roadblocks on DNA strands. This Review will illustrate how various cellular components including replication protein A (RPA), Bloom syndrome protein (BLM), and Pif1 helicases are capable of unfolding qDNA. Techniques such as single-molecule fluorescence resonance energy transfer (smFRET), often in conjunction with the aforementioned force-based techniques, have proven extremely effective at elucidating the factors underpinning the mechanisms by which these proteins unwind qDNA structures. We will provide insight into how single-molecule tools have facilitated the direct visualization of qDNA roadblocks and also showcase results obtained from experiments designed to examine the ability of G-quadruplexes to limit the access of specific cellular proteins normally associated with telomeres.

Bioactivated and PEG-Protected Circa 2nm Gold Nanoparticles for in Cell Labelling and Cryo-Electron Microscopy.

Advances in cryo-electron microscopy (EM) enable imaging of protein assemblies within mammalian cells in a near native state when samples are preserved by cryogenic vitrification. To accompany this progress, specialized EM labelling protocols must be developed. Gold nanoparticles (AuNPs) of 2nm are synthesized and functionalized to bind selected intracellular targets inside living human cells and to be detected in vitreous sections. As a proof of concept, thioaminobenzoate-, thionitrobenzoate-coordinated gold nanoparticles are functionalized on their surface with SV40 Nuclear Localization Signal (NLS)-containing peptides and 2kDa polyethyleneglycols (PEG) by thiolate exchange to target the importin-mediated nuclear machinery and facilitate cytosolic diffusion by shielding the AuNP surface from non-specific binding to cell components, respectively. After delivery by electroporation into the cytoplasm of living human cells, the PEG-coated AuNPs diffuse freely in the cytoplasm but do not enter the nucleus. Incorporation of NLS within the PEG coverage promotes a quick nuclear import of the nanoparticles in relation to the density of NLS onto the AuNPs. Cryo-EM of vitreous cell sections demonstrate the presence of 2nm AuNPs as single entities in the nucleus. Biofunctionalized AuNPs combined with live-cell electroporation procedures are thus potent labeling tools for the identification of macromolecules in cellular cryo-EM.

Reunderstand and Discuss the Hardness Limits of RCC-M M5110 Part Materials

RCC-M M5110 is the procurement code for rolling or forging bars for bolts and drive rods of class 1, 2 and 3 equipment of nuclear power plants. The individual high strength steels involved in this specification specify only the minimum hardness of the material and have no limit on the maximum hardness. The material selection of major components of nuclear power machinery equipment must meet the requirements of load bearing and safe operation, and ensure sufficient strength and hardness, which is especially important for the material selection of connecting bolts and driving rods of nuclear power equipment. As the strength and hardness of high strength steel increase, the plasticity and toughness of materials decrease sharply, as do the mechanical properties and corrosion resistance, increasing the risk of bolt fracture and equipment damage. In order to ensure the strength and hardness of metal materials, it is necessary to consider the comprehensive mechanical properties and corrosion resistance of the material, including resistance stress corrosion cracking (SCC). Improving strength and hardness should not be the only goal of improving the performance of metal materials. In the formulation and selection of raw material standards, the mechanical properties of important equipment materials, such as hardness, must be limited. To meet the requirements of "redundant design" and "defense in depth" of nuclear power equipment. This paper will discuss how to reunderstand the the hardness limits of RCC-M M 5110 materials.

Modulation of nuclear membrane repair machinery by nano-needle arrays

Abnormal nuclear structure and temporary nuclear membrane rupture have been identified by recent studies as common characteristics in cancer cells. However, cancer cell lines have been found to repeatedly proliferate after nuclear membrane rupture, implicating a mechanism of over-activated nuclear membrane repair outside of the normal cell cycle. Therefore, the modulation of nuclear membrane repair machinery has emerged as a new target for cancer therapeutics. The identification of therapeutic targets depends on the visualization, characterization and mechanistic understanding of repair machinery in health and disease, with single cell resolution. Current methods for investigating nuclear repair machinery suffer from limitations in spatial resolution, throughput, control and reproducibility of activated repair mechanisms. In this work, we propose a new platform consisting of arrays of nano-scale needles with varying geometries to induce sharp curvature and potentially rupture of the nuclear membrane. We used immunocytochemistry to visualize a nuclear DNA repair protein (KU80) that mislocalized to the cytoplasm as a marker of nuclear membrane rupture, and Lamin A/C to observe nuclear membrane shape on the nanoneedles. In comparison to a flat portion of the nanopillar array, the nuclear content of seeded cells are displaced spatially where nanostructures were present. Additionally, we observed frequent ruptures of the nuclear membrane of cells on nanoneedles. Our results imply that high curvature results in higher frequencies of nuclear membrane rupture, potentially activating repair machinery at the tips of nanoneedles. Thus, designing optimal geometries of nanoneedles can provide a platform with improved spatial resolution, throughput, control and reproducibility for inducing membrane rupture, and activating specific repair machinery. Ultimately, our platform can enable more efficient and controlled investigations into the nuclear membrane rupture and repair process, leading to new discoveries and research avenues for modulating this process in cancer.

A germline-targeted genetic screen for xrn-2 suppressors identifies a novel gene C34C12.2 in Caenorhabditis elegans.

XRN2 is an evolutionarily conserved 5'-to-3' exoribonuclease, which degrades or trims various types of RNA in the nucleus. Although XRN-2 is essential for embryogenesis, larval development and reproduction in Caenorhabditis elegans, relevant molecular pathways remain unidentified. Here we create a germline-specific xrn-2 conditional mutant and perform a mutagenesis screen for suppressors of sterility. Loss-of-function alleles of dpy-10, osr-1, ptr-6 and C34C12.2 genes are identified. Depletion of DPY-10, OSR-1 or PTR-6 increases expression of gpdh-1 that encodes a glycerol-3-phosphate dehydrogenase, thereby elevates glycerol accumulation to suppress sterility of the mutant. The C34C12.2 protein is predominantly localized in the nucleolus of germ cells and shows a similarity to Saccharomyces cerevisiae Net1, which is involved in rDNA silencing. Depletion of NRDE-2, a putative interacting partner of C34C12.2 and a component of the nuclear RNAi machinery, restores fertility to the xrn-2 conditional mutant. These results may help to identify an essential role of XRN-2 in germline development.

Tritiated Steel Micro-Particles: Computational Dosimetry and Prediction of Radiation-Induced DNA Damage for In Vitro Cell Culture Exposures.

Biological effects of radioactive particles can be experimentally investigated in vitro as a function of particle concentration, specific activity and exposure time. However, a careful dosimetric analysis is needed to elucidate the role of radiation emitted by radioactive products in inducing cyto- and geno-toxicity: the quantification of radiation dose is essential to eventually inform dose-risk correlations. This is even more fundamental when radioactive particles are short-range emitters and when they have a chemical speciation that might further concur to the heterogeneity of energy deposition at the cellular and sub-cellular level. To this aim, we need to use computational models. In this work, we made use of a Monte Carlo radiation transport code to perform a computational dosimetric reconstruction for in vitro exposure of cells to tritiated steel particles of micrometric size. Particles of this kind have been identified as worth of attention in nuclear power industry and research: tritium easily permeates in steel elements of nuclear reactor machinery, and mechanical operations on these elements (e.g., sawing) during decommissioning of old facilities can result in particle dispersion, leading to human exposure via inhalation. Considering the software replica of a representative in vitro setup to study the effect of such particles, we therefore modelled the radiation field due to the presence of particles in proximity of cells. We developed a computational approach to reconstruct the dose range to individual cell nuclei in contact with a particle, as well as the fraction of "hit" cells and the average dose for the whole cell population, as a function of particle concentration in the culture medium. The dosimetric analysis also provided the basis to make predictions on tritium-induced DNA damage: we estimated the dose-dependent expected yield of DNA double strand breaks due to tritiated steel particle radiation, as an indicator of their expected biological effectiveness.

Acetylation of Nup62 by TIP60 ensures accurate chromosome segregation in mitosis.

Stable transmission of genetic information during cell division requires faithful mitotic spindle assembly and chromosome segregation. In eukaryotic cells, nuclear envelope breakdown (NEBD) is required for proper chromosome segregation. Although a list of mitotic kinases has been implicated in NEBD, how they coordinate their activity to dissolve the nuclear envelope and protein machinery such as nuclear pore complexes was unclear. Here, we identified a regulatory mechanism in which Nup62 is acetylated by TIP60 in human cell division. Nup62 is a novel substrate of TIP60, and the acetylation of Lys432 by TIP60 dissolves nucleoporin Nup62-Nup58-Nup54 complex during entry into mitosis. Importantly, this acetylation-elicited remodeling of nucleoporin complex promotes the distribution of Nup62 to the mitotic spindle, which is indispensable for orchestrating correct spindle orientation. Moreover, suppression of Nup62 perturbs accurate chromosome segregation during mitosis. These results establish a previously uncharacterized regulatory mechanism in which TIP60-elicited nucleoporin dynamics promotes chromosome segregation in mitosis.

Nuclear entry and egress of parvoviruses.

Parvoviruses are small non-enveloped single-stranded DNA viruses, which depend on host cell nuclear transcriptional and replication machinery. After endosomal exposure of nuclear localization sequence and a phospholipase A2 domain on the capsid surface, and escape into the cytosol, parvovirus capsids enter the nucleus. Due to the small capsid diameter of 18-26 nm, intact capsids can potentially pass into the nucleus through nuclear pore complexes (NPCs). This might be facilitated by active nuclear import, but capsids may also follow an alternative entry pathway that includes activation of mitotic factors and local transient disruption of the nuclear envelope. The nuclear entry is followed by currently undefined events of viral genome uncoating. After genome release, viral replication compartments are initiated and infection proceeds. Parvoviral genomes replicate during cellular S phase followed by nuclear capsid assembly during virus-induced S/G2 cell cycle arrest. Nuclear egress of capsids occurs upon nuclear envelope degradation during apoptosis and cell lysis. An alternative pathway for nuclear export has been described using active transport through the NPC mediated by the chromosome region maintenance 1 protein, CRM1, which is enhanced by phosphorylation of the N-terminal domain of VP2. However, other alternative but not yet uncharacterized nuclear export pathways cannot be excluded.

Nab3 nuclear granule accumulation is driven by respiratory capacity.

Numerous biological processes involve proteins capable of transiently assembling into subcellular compartments necessary for cellular functions. One process is the RNA polymerase II transcription cycle which involves initiation, elongation, co-transcriptional modification of nascent RNA, and termination. The essential yeast transcription termination factor Nab3 is required for termination of small non-coding RNAs and accumulates into a compact nuclear granule upon glucose removal. Nab3 nuclear granule accumulation varies in penetrance across yeast strains and a higher Nab3 granule accumulation phenotype is associated with petite strains, suggesting a possible ATP-dependent mechanism for granule disassembly. Here, we demonstrate the uncoupling of mitochondrial oxidative phosphorylation by drug treatment or deletions of nuclear-encoded ATP synthase subunit genes were sufficient to increase Nab3 granule accumulation and led to an inability to proliferate during prolonged glucose deprivation, which requires respiration. Additionally, by enriching for respiration competent cells from a petite-prone strain, we generated a low granule-accumulating strain from a relatively high one, providing another link between respiratory competency and Nab3 granules. Consistent with the resulting idea that ATP is involved in granule accumulation, the addition of extracellular ATP to semi-permeabilized cells was sufficient to reduce Nab3 granule accumulation. Deleting the SKY1 gene, which encodes a kinase that phosphorylates nuclear SR repeat-containing proteins and is involved in efficient stress granule disassembly, also resulted in increased granule accumulation. This observation implicates Sky1 in Nab3 granule biogenesis. Taken together, these findings suggest there is normally an equilibrium between termination factor granule assembly and disassembly mediated by ATP-requiring nuclear machinery.

Chromatin Dynamics in Digestive System Cancer: Commander and Regulator.

Digestive system tumors have a poor prognosis due to complex anatomy, insidious onset, challenges in early diagnosis, and chemoresistance. Epidemiological statistics has verified that digestive system tumors rank first in tumor-related death. Although a great number of studies are devoted to the molecular biological mechanism, early diagnostic markers, and application of new targeted drugs in digestive system tumors, the therapeutic effect is still not satisfactory. Epigenomic alterations including histone modification and chromatin remodeling are present in human cancers and are now known to cooperate with genetic changes to drive the cancer phenotype. Chromatin is the carrier of genetic information and consists of DNA, histones, non-histone proteins, and a small amount of RNA. Chromatin and nucleosomes control the stability of the eukaryotic genome and regulate DNA processes such as transcription, replication, and repair. The dynamic structure of chromatin plays a key role in this regulatory function. Structural fluctuations expose internal DNA and thus provide access to the nuclear machinery. The dynamic changes are affected by various complexes and epigenetic modifications. Variation of chromatin dynamics produces early and superior regulation of the expression of related genes and downstream pathways, thereby controlling tumor development. Intervention at the chromatin level can change the process of cancer earlier and is a feasible option for future tumor diagnosis and treatment. In this review, we introduced chromatin dynamics including chromatin remodeling, histone modifications, and chromatin accessibility, and current research on chromatin regulation in digestive system tumors was also summarized.

Remaining Useful Life Prediction of Nuclear Power Machinery Based on an Exponential Degradation Model

Aiming at solving the problems of small fault data samples and insufficient remaining useful life (RUL) prediction accuracy of nuclear power machinery, a method based on an exponential degradation model is proposed to predict the RUL of equipment after the failure warning system alarm. After data preprocessing, time-domain feature extraction, selection, and dimensionality reduction fusion of multiple degradation variables, the exponential degradation model is constructed based on the Bayesian process, and prior information is used. As an application, the RUL of a nuclear power turbine was calculated based on actual monitoring data, the α − λ precision curve was used to evaluate the prediction effect, and the RUL prediction results verified the effectiveness of the proposed method.

Conserved Apical Proline Regulating the Structure and DNA Binding Properties of Helicobacter pylori Histone-like DNA Binding Protein (Hup).

Prokaryotic cells lack a proper dedicated nuclear arrangement machinery. A set of proteins known as nucleoid associated proteins (NAPs) perform opening and closure of nucleic acids, behest cellular requirement. Among these, a special class of proteins analogous to eukaryotic histones popularly known as histone-like (HU) DNA binding proteins facilitate the nucleic acid folding/compaction thereby regulating gene architecture and gene regulation. DNA compaction and DNA protection in Helicobacter pylori is performed by HU protein (Hup). To dissect and galvanize the role of proline residue in the binding of Hup with DNA, the structure-dynamics-functional relationship of Hup-P64A variant was analyzed. NMR and biophysical studies evidenced that Hup-P64A protein attenuated DNA-binding and induced structural/stability changes in the DNA binding domain (DBD). Moreover, molecular dynamics simulations and 15N relaxation studies established the reduced conformational dynamics of P64A protein. This comprehensive study dissected the exclusive role of evolutionarily conserved apical proline residue in regulating the structure and DNA binding of Hup protein as P64 is presumed to be involved in the external leverage mechanism responsible for DNA bending and packaging, as proline rings wedge into the DNA backbone through intercalation besides their significant role in DNA binding.

Advances in nuclear power system design and fault-based condition monitoring towards safety of nuclear-powered ships

The increasing need for cleaner and sustainable shipping operations, especially in the Arctic waters, has informed the rising technological advancements in ship nuclear power machinery (SNPM) design, such as the generation IV (GenIV) small modular reactor (SMR). Nuclear technology has also provided a potential alternative to decarbonization in the maritime industry. The recent design advances present safety challenges that need to be comprehensively studied and understood. The current study shows a review of the advancements in nuclear plant design and fault-based analysis towards safety in maritime operations. A systematic approach is adopted to explore the design improvements, operational challenges, fault diagnostics methodologies, and risk assessment frameworks for SNPM. While exploring the current state of knowledge on system design and fault-based risk assessment, the study highlights key research opportunities to provide a robust risk management framework towards safer nuclear-powered shipping operations, such as digitalization, resilient system design, and the internet of things (IoT). This study finds that the state of knowledge is inexhaustive and proposes a preliminary hybrid framework, able to integrate the hierarchical hazard structure into a fault-based risk aggregation methodology for risk-informed decision making in nuclear-powered shipping operations.

An approach for obtaining surface residual stress based on indentation test and strain measurement

Abstract During the welding process of welded joints in nuclear power plants, uneven deformation caused by temperature changes will lead to the generation of residual stress. Aiming at the problem that the demarcate experiment is easy to cause errors when the traditional indentation strain method is used to measure the residual stress. Using 316L austenitic stainless steel as an example, ABAQUS finite element simulation is combined with physical experiments to study the effects of different indentation depths on the strain increment after unloading under a state of residual stress. The results show that the indentation depth and strain increment are linearly related. Based on the linear relationship, an independent parameter that characterizes the residual stress is obtained and a model between the parameter and residual stress established. It provides a certain basis for the convenience and precise detection of residual stress in nuclear power plants machinery structures in service.

Nuclear mRNA Quality Control and Cytoplasmic NMD Are Linked by the Guard Proteins Gbp2 and Hrb1.

Pre-mRNA splicing is critical for cells, as defects in this process can lead to altered open reading frames and defective proteins, potentially causing neurodegenerative diseases and cancer. Introns are removed in the nucleus and splicing is documented by the addition of exon-junction-complexes (EJCs) at exon-exon boundaries. This “memory” of splicing events is important for the ribosome, which translates the RNAs in the cytoplasm. In case a stop codon was detected before an EJC, translation is blocked and the RNA is eliminated by the nonsense-mediated decay (NMD). In the model organism Saccharomyces cerevisiae, two guard proteins, Gbp2 and Hrb1, have been identified as nuclear quality control factors for splicing. In their absence, intron-containing mRNAs leak into the cytoplasm. Their presence retains transcripts until the process is completed and they release the mRNAs by recruitment of the export factor Mex67. On transcripts that experience splicing problems, these guard proteins recruit the nuclear RNA degradation machinery. Interestingly, they continue their quality control function on exported transcripts. They support NMD by inhibiting translation and recruiting the cytoplasmic degradation factors. In this way, they link the nuclear and cytoplasmic quality control systems. These discoveries are also intriguing for humans, as homologues of these guard proteins are present also in multicellular organisms. Here, we provide an overview of the quality control mechanisms of pre-mRNA splicing, and present Gbp2 and Hrb1, as well as their human counterparts, as important players in these pathways.