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When single-phase ground faults occur in distribution systems, the fault characteristics of zero-sequence current signals are not prominent. They are quickly submerged in noise, leading to difficulties in fault section location. This paper proposes a method for fault section location in distribution networks based on improved empirical wavelet transform (IEWT) and GINs to address this issue. Firstly, based on kurtosis, EWT is optimized using the N-point search method to decompose the zero-sequence current signal into modal components. Noise is filtered out through weighted permutation entropy (WPE), and signal reconstruction is performed to obtain the denoised zero-sequence current signal. Subsequently, GINs are employed for graph classification tasks. According to the topology of the distribution network, the corresponding graph is constructed as the input to the GIN. The denoised zero-sequence current signal is the node input for the GIN. The GIN autonomously explores the features of each graph structure to achieve fault section location. The experimental results demonstrate that this method has strong noise resistance, with a fault section location accuracy of up to 99.95%, effectively completing fault section location in distribution networks. [ABSTRACT FROM AUTHOR]

Abstract Gastrointestinal nematodes (GINs) are major constraints to health and productivity of small ruminants. Methods of their control relies mainly on anthelmintic drugs; however, the indiscriminate use of these drugs could lead to the development of anthelmintic resistance (AR). This study aimed to investigate the epidemiology of GINs infection, and field evaluation of anthelmintic efficacy in sheep. The epidemiological data were collected using a cross-sectional study design while a farm-based field study design was employed for the evaluation of anthelminthic efficacy. Furthermore, standard parasitological techniques were employed for qualitative and quantitative worm identification. The overall prevalence indicated 50.3%. Six genera of GINs (Haemonchus, Trichostrongylus, Oesophagostomum/Chabertia, Trichuris, Teladosargia/Ostertagia and Nematodirus) were identified. Among the identified genera, Haemonchus (25.4%) and Trichostrongylus (24.8%) were the dominant genera followed by mixed infection (21.8%), Oesophagostomum/Chabertia (10.4%), Trichuris (7.8%), Teladosargia (Ostertagia) (5.7%) and Nematodirus (4.1%). Mixed infections consisted either of double infections with Haemonchus and Trichostrongylus, or triple infections with Haemonchus, Trichostrongylus and Trichuris. The McMaster egg counting results showed that the mean EPG of infected sheep was 845.6. The results also showed 66 (34.2%), 101 (52.3%) and 26 (13.5%) sheep had low, moderate and heavy worm burden, respectively. Albendazole and Ivermectin showed low efficacy (percentage reductions = 90% and 92%; 95% lower confidence limit = 82.1% and 83.6% respectively) whereas Tetramisole was effective (FECR% = 96.8%; 95% LCL = 93.4%). Factors such as age, body condition, management system and past deworming history of sheep were found to have a statistically significant (p

When single-phase ground faults occur in distribution systems, the fault characteristics of zero-sequence current signals are not prominent. They are quickly submerged in noise, leading to difficulties in fault section location. This paper proposes a method for fault section location in distribution networks based on improved empirical wavelet transform (IEWT) and GINs to address this issue. Firstly, based on kurtosis, EWT is optimized using the N-point search method to decompose the zero-sequence current signal into modal components. Noise is filtered out through weighted permutation entropy (WPE), and signal reconstruction is performed to obtain the denoised zero-sequence current signal. Subsequently, GINs are employed for graph classification tasks. According to the topology of the distribution network, the corresponding graph is constructed as the input to the GIN. The denoised zero-sequence current signal is the node input for the GIN. The GIN autonomously explores the features of each graph structure to achieve fault section location. The experimental results demonstrate that this method has strong noise resistance, with a fault section location accuracy of up to 99.95%, effectively completing fault section location in distribution networks.

ABSTRACT The identification of microbial genes essential for survival as those with lethal knockout phenotype (LKP) is a common strategy for functional interrogation of genomes. However, interpretation of the LKP is complicated because a substantial fraction of the genes with this phenotype remains poorly functionally characterized. Furthermore, many genes can exhibit LKP not because their products perform essential cellular functions but because their knockout activates the toxicity of other genes (conditionally essential genes). We analyzed the sets of LKP genes for two archaea, Methanococcus maripaludis and Sulfolobus islandicus, using a variety of computational approaches aiming to differentiate between essential and conditionally essential genes and to predict at least a general function for as many of the proteins encoded by these genes as possible. This analysis allowed us to predict the functions of several LKP genes including previously uncharacterized subunit of the GINS protein complex with an essential function in genome replication and of the KEOPS complex that is responsible for an essential tRNA modification as well as GRP protease implicated in protein quality control. Additionally, several novel antitoxins (conditionally essential genes) were predicted, and this prediction was experimentally validated by showing that the deletion of these genes together with the adjacent genes apparently encoding the cognate toxins caused no growth defect. We applied principal component analysis based on sequence and comparative genomic features showing that this approach can separate essential genes from conditionally essential ones and used it to predict essential genes in other archaeal genomes.IMPORTANCEOnly a relatively small fraction of the genes in any bacterium or archaeon is essential for survival as demonstrated by the lethal effect of their disruption. The identification of essential genes and their functions is crucial for understanding fundamental cell biology. However, many of the genes with a lethal knockout phenotype remain poorly functionally characterized, and furthermore, many genes can exhibit this phenotype not because their products perform essential cellular functions but because their knockout activates the toxicity of other genes. We applied state-of-the-art computational methods to predict the functions of a number of uncharacterized genes with the lethal knockout phenotype in two archaeal species and developed a computational approach to predict genes involved in essential functions. These findings advance the current understanding of key functionalities of archaeal cells.

يستعرض البحث بإيجاز تاريخ وتطور شرکات الحلج وکبس القطن في مصر وما يتبعها من أنشطة في الفترة من عام 1894 م إلي 1952م. حيث تأسست في مصر الکثير من شرکات حلج وکبس وتکرير القطن وجاء تأسيسها ضرورة ملحة لتواکب زراعة القطن التي بدأت زراعتها في عصر الوالي محمد علي باشا، والمحالج کانت ضرورية لحلج القطن بفصل بذرته عن شعيرات القطن، وما يتبع ذلک من تکرير للقطن وکبسه في بالات معده للشحن لاستخدامها في مصانع الغزل والنسيج وغير ذلک من استخدامات القطن المتعددة، واستخدامات بذرته في عصر الزيوت وغيرها من صناعات کانت بداية هذه المحالج بمحالج أربعة أنشأها محمد علي باشا کان أولها في القناطر الخيرية ثم اتبعها بثلاث أخرى لتتوالى بعد ذلک زيادة أعدادها مع زيادة زراعة مساحة القطن والطلب عليه، وجاءت زيادة أعداد المحالج بعد الاحتلال الإنجليزي وما يتبع ذلک من أنشطة ملحة بها وحتى عام 1952م. أقيمت محالج القطن في أکثرها بالتوازي مع إنشاء وبداية صناعة الغزل والنسيج التي أخذت في التطور من أنوال يدوية إلى مغازل وأنوال ميکانيکية، وهو نفس التطور الذي واکب صناعة حلج القطن وکبسه فقد کان حلج القطن في بدايته يدويا في عملية شاقة استهلکت الکثير من الإيدي العاملة والوقت والجهد، وأدخلت عليها التحسينات وأعمال التطور بصناعة دواليب الحلج التي استخدمت الثيران في تشغيلها ودورانها إلى أن استخدمت المحالج المطورة والتي أصبحت تدار بالديزل والکهرباء حتي عام 1952م. تتناول هذه الدراسة عدد من المحاور الرئيسية تتمثل في دعامات صناعة حلج وکبس القطن والتي لولاها لما کانت لها قائمة وانتشرت في مصر وأيضا في العالم، تأسيس شرکات محالج القطن وکبسه والتي تم انشائها خلال فترة الدراسة، أهم الأنشطة التابعه لتلک المحالج، مساهمة رؤوس أموال هذه الشرکات في دعم الاقتصاد المصري. Abstract The research briefly reviews the history and development of cotton gin and pressing companies in of cotton companies were established in Egypt, and its establishment came as an urgent Egypt and their activities in the period from 1894 to 1952. Many of the ginning, pressing and refining necessity to keep pace with the cultivation of cotton that began in the era of the Governor Muhammad Ali Pasha. Gins were necessary for scooping the cotton by separating its seed from its bristles, and consequently, for the subsequent refining of the cotton and compressing it in batches intended for shipment, for use in the textile factories and other multiple uses of cotton. The uses of cotton seeds in getting oils and other industries were the beginning of these gins, which started with four gins established by Muhammad Ali Pasha. The first of these was held in Al-Qanater-al-Khayriyah and then followed by three others before they were successively increased simultaneously with the increasing of the cotton cultivated area and the increase in the demand of Egyptian cotton. The increase in the number of gins occurred after the British occupation and the urgent activities that followed it until 1952. Cotton gins were set up in parallel with the establishment and beginning of the textile industry, that began to evolve from manual looms to spindles and mechanical looms; the same development that accompanied the ginning and compressing of cotton. The ginning of cotton started initially in a laborious process that consumed a lot of labor, time and effort. This was accompanied with improvements and developments in the manufacture of ginning racks that used bulls in operating and rotating them. This lasted until developed gin was used; a gin which became powered by diesel and electricity up to 1952. This study addresses a number of the main axes represented in the pillars of cotton ginning and pressing industry, without which it would not have existed and spread in Egypt, especially, and in the world, generally. In addition, it deals with establishing ginning and pressing cotton companies that were established during the period covered by the study. The study also tackles the most important activities of these gins, and finally, the contribution of these capital Companies in supporting the Egyptian economy.

Background: The GINS complex, composed of GINS1/2/3/4 subunits, is an essential structure of Cdc45-MCM-GINS (CMG) helicase and plays a vital role in establishing the DNA replication fork and chromosome replication. Meanwhile, GINS genes have been associated with the poor prognosis of various malignancies. However, the abnormal expression of GINS genes and their diagnostic and prognostic value in sarcomas (SARC) remain unclear.Methods: Oncomine, Gene Expression Profiling Interactive Analysis (GEPIA), Kaplan-Meier Plotter, Cancer cell line encyclopedia (CCLE), The University of Alabama at Birmingham Cancer Data Analysis Portal (UALCAN), R studio, and Tumor Immune Estimation Resource (TIMER) were used to analyze the expression profiles, prognostic value, biological function, ceRNA, and immune infiltration associated with GINS genes in sarcomas.Results: We found that GINS1/2/3/4 genes exhibited significantly upregulated transcription levels in SARC samples compared to non-tumor tissues and exhibited high expression levels in sarcoma cell lines. In addition, SARC patients with increased expression levels of GINS1/2/3/4 showed poorer survival rates. Immune infiltration analysis showed that GINS subunits were closely associated with the infiltration of immune cells in sarcomas.Conclusion: Our research identified GINS subunits as potential diagnostic and prognostic biological targets in SARC and elucidated their underlying effects in the genesis and progression of SARC. These results may provide new opportunities and research directions for targeted sarcoma therapy.

Metazoan genomes are copied bidirectionally from thousands of replication origins. Replication initiation entails the assembly and activation of two CMG helicases (Cdc45⋅Mcm2-7⋅GINS) at each origin. This requires several replication firing factors (including TopBP1, RecQL4, and DONSON) whose exact roles are still under debate. How two helicases are correctly assembled and activated at each origin is a long-standing question. By visualizing the recruitment of GINS, Cdc45, TopBP1, RecQL4, and DONSON in real time, we uncovered that replication initiation is surprisingly dynamic. First, TopBP1 transiently binds to the origin and dissociates before the start of DNA synthesis. Second, two Cdc45 are recruited together, even though Cdc45 alone cannot dimerize. Next, two copies of DONSON and two GINS simultaneously arrive at the origin, completing the assembly of two CMG helicases. Finally, RecQL4 is recruited to the CMG⋅DONSON⋅DONSON⋅CMG complex and promotes DONSON dissociation and CMG activation via its ATPase activity. [Display omitted] • Single-molecule experiments shed light on how DNA replication is initiated • Reveals the order in which replication initiation factors are recruited to origins • 2-fold symmetry is established by dimerization scaffolds that recruit GINS and Cdc45 • The ATPase activity of RecQL4 plays an important role in efficient origin firing Single-molecule imaging was used to visualize how DNA replication initiates bidirectionally from individual origins, providing critical insights into the function and dynamics of several key replication initiation factors. [ABSTRACT FROM AUTHOR]

Replisomes follow a schedule in which replication of DNA in euchromatin is early in S phase while sequences in heterochromatin replicate late. Impediments to DNA replication, referred to as replication stress, can stall replication forks triggering activation of the ATR kinase and downstream pathways. While there is substantial literature on the local consequences of replisome stalling–double strand breaks, reversed forks, or genomic rearrangements–there is limited understanding of the determinants of replisome stalling vs. continued progression. Although many proteins are recruited to stalled replisomes, current models assume a single species of “stressed” replisome, independent of genomic location. Here we describe our approach to visualizing replication fork encounters with the potent block imposed by a DNA interstrand crosslink (ICL) and our discovery of an unexpected pathway of replication restart (traverse) past an intact ICL. Additionally, we found two biochemically distinct replisomes distinguished by activity in different stages of S phase and chromatin environment. Each contains different proteins that contribute to ICL traverse.

Bacterial RecJ exhibits 5'→3' exonuclease activity that is specific to ssDNA; however, archaeal RecJs show 5' or 3' exonuclease activity. The hyperthermophilic archaea Methanocaldococcus jannaschii encodes the 5'-exonuclease MjRecJ1 and the 3'-exonuclease MjRecJ2. In addition to nuclease activity, archaeal RecJ interacts with GINS, a structural subcomplex of the replicative DNA helicase complex. However, MjRecJ1 and MjRecJ2 do not interact with MjGINS. Here, we report the structural basis for the inability of the MjRecJ2 homologous dimer to interact with MjGINS and its efficient 3' hydrolysis polarity for short dinucleotides. Based on the crystal structure of MjRecJ2, we propose that the interaction surface of the MjRecJ2 dimer overlaps the potential interaction surface for MjGINS and blocks the formation of the MjRecJ2-GINS complex. Exposing the interaction surface of the MjRecJ2 dimer restores its interaction with MjGINS. The cocrystal structures of MjRecJ2 with substrate dideoxynucleotides or product dCMP/CMP show that MjRecJ2 has a short substrate binding patch, which is perpendicular to the longer patch of bacterial RecJ. Our results provide new insights into the function and diversification of archaeal RecJ/Cdc45 proteins., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Goats greatly influence the economic sustainability of rural communities. However, parasitic diseases, especially gastrointestinal nematodes (GINs) are a major constraint on profitable small ruminants' production worldwide. During July- 2015 to June- 2016, we conducted a cross sectional study within seven topographic zones of Bangladesh to explore the level of infection and associated risk factors of GINs infections of goats. The study followed standard flotation and modified McMaster techniques. Among 1998 samples from goats; 1241 (62.1%) were found to be infected with one or more species of GINs by fecal examination for nematode eggs. The identified nematodes were strongyles (51.9%), Strongyloides sp. (19.0%) and Trichuris spp. (2.9%). By coproculture, we identified Haemonchus spp., Oesophagostomum spp., Trichostrongylus spp. and Bunostomum spp. in the different topographic zones. According to univariate analysis; young age, other breed than Black Bengal, animals in poor condition, backyard rearing system, muddy housing, illiterate farmers and rainy season were found significantly associated with GINs infections. Besides, other breed than Black Bengal, animals in poor condition, backyard rearing system, muddy housing and illiterate farmers were identified as the risk factors of GINs infections in goats. This is the first detailed epidemiological investigation of GINs of goats in Bangladesh. The epidemiological findings are expected to help formulate effective control strategies against GINs infections in goats by improving health status of animals, management system and education level of the farmers.

Abstract The eukaryotic GINS, Cdc45, and minichromosome maintenance proteins form an essential complex that moves with the DNA replication fork. The GINS protein complex has also been reported to associate with DNA polymerase. In archaea, the third domain of life, DNA polymerase D (PolD) is essential for DNA replication, and the genes encoding PolDs exist only in the genomes of archaea. The archaeal GAN (GINS‐associated nuclease) is believed to be a homolog of the eukaryotic Cdc45. In this study, we found that the Thermococcus sp. 4557 DP1 (small subunit of PolD) interacted with GINS15 in vitro, and the 3′–5′ exonuclease activity of DP1 was inhibited by GINS15. We also demonstrated that the GAN, GINS15, and DP1 proteins interact to form a complex adapting a GAN–GINS15–DP1 order. The results of this study imply that the complex constitutes a core of the DNA replisome in archaea.

The identification of microbial genes essential for survival as those with lethal knockout phenotype (LKP) is a common strategy for functional interrogation of genomes. However, interpretation of the LKP is complicated because a substantial fraction of the genes with this phenotype remains poorly functionally characterized. Furthermore, many genes can exhibit LKP not because their products perform essential cellular functions but because their knockout activates the toxicity of other genes (conditionally essential genes). We analyzed the sets of LKP genes for two archaea, Methanococcus maripaludis and Sulfolobus islandicus, using a variety of computational approaches aiming to differentiate between essential and conditionally essential genes and to predict at least a general function for as many of the proteins encoded by these genes as possible. This analysis allowed us to predict the functions of several LKP genes including previously uncharacterized subunit of the GINS protein complex with an essential function in genome replication and of the KEOPS complex that is responsible for an essential tRNA modification as well as GRP protease implicated in protein quality control. Additionally, several novel antitoxins (conditionally essential genes) were predicted, and this prediction was experimentally validated by showing that the deletion of these genes together with the adjacent genes apparently encoding the cognate toxins caused no growth defect. We applied principal component analysis based on sequence and comparative genomic features showing that this approach can separate essential genes from conditionally essential ones and used it to predict essential genes in other archaeal genomes.IMPORTANCEOnly a relatively small fraction of the genes in any bacterium or archaeon is essential for survival as demonstrated by the lethal effect of their disruption. The identification of essential genes and their functions is crucial for understanding fundamental cell biology. However, many of the genes with a lethal knockout phenotype remain poorly functionally characterized, and furthermore, many genes can exhibit this phenotype not because their products perform essential cellular functions but because their knockout activates the toxicity of other genes. We applied state-of-the-art computational methods to predict the functions of a number of uncharacterized genes with the lethal knockout phenotype in two archaeal species and developed a computational approach to predict genes involved in essential functions. These findings advance the current understanding of key functionalities of archaeal cells., Competing Interests: The authors declare no conflict of interest.

During cell division, the genome of each eukaryotic cell is copied by thousands of replisomes-large protein complexes consisting of several dozen proteins. Recent studies suggest that the eukaryotic replisome is much more dynamic than previously thought. To directly visualize replisome dynamics in a physiological context, we recently developed a single-molecule approach for imaging replication proteins in Xenopus egg extracts. These extracts contain all the soluble nuclear proteins and faithfully recapitulate DNA replication and repair in vitro, serving as a powerful platform for studying the mechanisms of genome maintenance. Here we present detailed protocols for conducting single-molecule experiments in nuclear egg extracts and preparing key reagents. This workflow can be easily adapted to visualize the dynamics and function of other proteins implicated in DNA replication and repair., Competing Interests: Declaration of interests Authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.)

Gastrointestinal nematodes (GINs) of ruminants are prevalent and have major economic impacts worldwide. The insight studies of immune responses triggered against GINs are of great concern to understand interaction between host's immune system and parasite. T-helper 2 cytokines drive the effector cell mechanisms which include eosinophils and mast cells. The immune responses are controlled by Th2 secreted interleukins (IL); IL3, IL-4, IL-5, IL-9, IL-10 and IL-13. B-Cell immune response is incorporated in defense mechanisms developed against GINs specially immunoglobulins (Ig); IgA, IgE and IgG. The immune resistance of the infected host is presented by failure of larval establishment or hypobiosis, low worm burden and decreased female fecundity. The host-parasite interaction is a complex series that affected by host's genetic constitution, nutrition, age and physiological status. The GINs have different immune evasion mechanisms to improve their survival within the host. Also, management of the host influences GINs parasitism. Thus, the aim of this review is to highlight the hallmarks of immune responses that endorse GINs parasitism. The insights studies of the triggered immune responses developed against GINs will improve the appropriate protective immune strategy. [ABSTRACT FROM AUTHOR]

The migration of chromosomes during mitosis is mediated primarily by kinesins that bind to the chromosomes and move along the microtubules, exerting pulling and pushing forces on the centrosomes. We report that a DNA replication protein, Sld5, localizes to the centrosomes, resisting the microtubular pulling forces experienced during chromosome congression. In the absence of Sld5, centriolar satellites, which normally cluster around the centrosomes, are dissipated throughout the cytoplasm, resulting in the loss of their known function of recruiting the centrosomal protein, pericentrin. We observed that Sld5-deficient centrosomes lacking pericentrin were unable to endure the CENP-E- and Kid-mediated microtubular forces that converge on the centrosomes during chromosome congression, resulting in monocentriolar and acentriolar spindle poles. The minus-end-directed kinesin-14 motor protein, HSET, sustains the traction forces that mediate centrosomal fragmentation in Sld5-depleted cells. Thus, we report that a DNA replication protein has an as yet unknown function of ensuring spindle pole resistance to traction forces exerted during chromosome congression. [ABSTRACT FROM AUTHOR]

The implementation into the GINS CNES geodetic software of a more efficient filter was needed to satisfy the users who wanted to compute high-rate GNSS PPP solutions. We selected the SRI approach and a QR factorization technique including an innovative algorithm which optimizes the matrix reduction step. A full description of this algorithm is given for future users. The new capacities of the software have been tested using a set of 1 Hz data from the Japanese GEONET network including the Mw 9.0 2011 Tohoku earthquake. Station coordinates solution agreed at a sub-decimeter level with previous publications as well as with solutions we computed with the National Resource Canada software. An additional benefit from the implementation of the SRI filter is the capability to estimate high-rate tropospheric parameters too. As the CPU time to estimate a 1 Hz kinematic solution from 1 h of data is now less than 1 min we could produced series of coordinates for the full 1300 stations of the Japanese network. The corresponding movie shows the impressive co-seismic deformation as well as the wave propagation along the island. The processing was straightforward using a cluster of PCs which illustrates the new potentiality of the GINS software for massive network high rate PPP processing. [ABSTRACT FROM AUTHOR]

Bacterial nuclease RecJ, which exists in almost all bacterial species, specifically degrades single-stranded (ss) DNA in the 5' to 3' direction. Some archaeal phyla, except Crenarchaea, also encode RecJ homologs. Compared with bacterial RecJ, archaeal RecJ exhibits a largely different amino acid sequence and domain organization. Archaeal RecJs from Thermococcus kodakarensis and Pyrococcus furiosus show 5'→3' exonuclease activity on ssDNA. Interestingly, more than one RecJ exists in some Euryarchaeota classes, such as Methanomicrobia, Methanococci, Methanomicrobia, Methanobacteria, and Archaeoglobi. Here we report the biochemical characterization of two RecJs from Methanocaldococcus jannaschii, the long RecJ1 (MJ0977) and short RecJ2 (MJ0831) to understand their enzymatic properties. RecJ1 is a 5'→3' exonuclease with a preference to ssDNA; however, RecJ2 is a 3'→5' exonuclease with a preference to ssRNA. The 5' terminal phosphate promotes RecJ1 activity, but the 3' terminal phosphate inhibits RecJ2 nuclease. Go-Ichi-Ni-San (GINS) complex does not interact with two RecJs and does not promote their nuclease activities. Finally, we discuss the diversity, function, and molecular evolution of RecJ in archaeal taxonomy. Our analyses provide insight into the function and evolution of conserved archaeal RecJ/eukaryotic Cdc45 protein. [ABSTRACT FROM AUTHOR]

The replicative helicase unwinds parental double-stranded DNA at a replication fork to provide single-stranded DNA templates for the replicative polymerases. In eukaryotes, the replicative helicase is composed of the Cdc45 protein, the heterohexameric ring-shaped Mcm2-7 complex and the tetrameric GINS complex (CMG). The CMG proteins bind directly to DNA as demonstrated by experiments with purified proteins. The mechanism and function of these DNA-protein interactions are presently being investigated and a number of important discoveries relating to how the helicase proteins interact with DNA have been reported recently. While some of the protein-DNA interactions directly relate to the unwinding function of the enzyme complex, other protein-DNA interactions may be important for minichromosome maintenance (MCM) loading, origin melting or replication stress. This review describes our current understanding of how the eukaryotic replicative helicase subunits interact with DNA structures in vitro and proposed models for the in vivo functions of replicative helicase-DNA interactions are also described. [ABSTRACT FROM AUTHOR]

In eukaryotes, proper loading and activation of MCM helicase at chromosomal origins plays a central role in DNA replication. Activation of MCM helicase requires its association with CDC45-GINS complex, but the mechanism of how this complex activates MCM helicase is poorly understood. Here we identified SIK1 (salt-inducible kinase 1), an AMPK related protein kinase, as a molecular link that connects GINS complex with MCM helicase activity. We demonstrated that Sld5 a component of GINS complex interacts with SIK1 and recruits it to the sites of DNA replication at the onset of S phase. Depletion of SIK1 leads to defective DNA replication. Further, we showed that SIK1 phosphorylates MCM2 at five conserved residues at its N-terminus, which is essential for the activation of MCM helicase. Collectively, our results suggest SIK1 as a novel integral component of CMG replicative helicase during eukaryotic DNA replication. [ABSTRACT FROM AUTHOR]

GINS2, a subunit of GINS complex, is critical for the initiation of DNA replication and DNA replication fork progression. The expression of GINS2 is misregulated in many malignant tumors, such as leukemia, breast cancer and melanoma. However, the role of GINS in breast cancer remains poorly characterized. We investigate the possible effect and particular mechanism of GINS in breast cancer cells. We showed that expression of GINS2 is enriched in triple negative breast cancer (TNBC) cell lines. Furthermore, GINS2 knockdown decreased the growth, invasive ability and stem-like property of TNBC cells. Mechanistically, silencing of GINS2 in TNBC cells caused dramatic decrease of matrix metalloproteinase-9 (MMP9). Finally, the abundance of GINS2 correlated with the advance stages of tumor in human TNBC patients. Our studies provided insight into the molecular regulation of TNBC progression and invasion. More importantly, our data suggest that GINS2 could be an outstanding therapeutic target for inhibiting invasive TNBC growth and metastasis. [ABSTRACT FROM AUTHOR]