Nuclear DNA

About: Nuclear DNA is a research topic. Over the lifetime, 3933 publications have been published within this topic receiving 185830 citations.

Non-linear accumulation of 8-hydroxy-2'-deoxyguanosine, a marker of oxidized DNA damage, during aging.

Abstract: Damage to DNA seems to be involved in aging and the etiology of age-associated degenerative diseases. The purpose of this study is to examine changes in DNA damage during aging. An oxidized nucleoside, 8-hydroxy-2'-deoxyguanosine (8-OHdG), is a proposed biomarker for DNA damaged by oxidative stress. The content of 8-OHdG in nuclear DNA isolated from brain, heart, liver, and kidneys of male Fischer 344 rats of different ages was measured, 8-OHdG can be detected selectively and sensitively at the fmol level by high performance liquid chromatography-electrochemical detection at an applied potential of +350 mV. The amount of 8-OHdG, expressed as the ratio to deoxyguanosine in nuclear DNA, in heart, liver, and kidney remained steady from 2 to 24 months and then increased progressively. The content of 8-OHdG in the DNA in brain showed no changes from 2 to 27 months, but was significantly higher in 30 month-old rats. There was a significant 2-fold increase in the amount of 8-OHdG in the nuclear DNA of all organs tested in 30 month-old rats as compared to 2-24 month-old rats. These results indicate that the accumulation of 8-OHdG in the DNA of rat organs begins at ages above 24 months.

Mitochondrial genetic medicine.

Abstract: Inherited mitochondrial DNA (mtDNA) diseases were discovered 30 years ago, and their characterization has provided a new perspective on the etiology of the common metabolic and degenerative diseases, cancer, and aging. The maternally inherited mtDNA contains 37 critical bioenergetic genes that are present in hundreds of copies per cell, but the 'mitochondrial genome' encompasses an additional 1,000-2,000 nuclear DNA (nDNA) mitochondrial genes. The interaction between these two mitochondrial genetic systems provides explanations for phenomena such as the non-Mendelian transmission of the common 'complex' diseases, age-related disease risk and progression, variable penetrance and expressivity, and gene-environment interactions. Thus, mtDNA genetics contributes to the quantitative and environmental components of human genetics that cannot be explained by Mendelian genetics. Because mtDNA is maternally inherited and cytoplasmic, it has fostered the first germline gene therapy, nuclear transplantation. However, effective interventions are still lacking for existing patients with mitochondrial dysfunction.

Mutations in the complex I NDUFS2 gene of patients with cardiomyopathy and encephalomyopathy.

Abstract: Human complex I is built up and regulated by genes encoded by the mitochondrial DNA (mtDNA) as well as the nuclear DNA (nDNA). In recent years, attention mainly focused on the relation between complex I deficiency and mtDNA mutations. However, a high percentage of consanguinity and an autosomal-recessive mode of inheritance observed within our patient group as well as the absence of common mtDNA mutations make a nuclear genetic cause likely. The NDUFS2 protein is part of complex I of many pro- and eukaryotes. The nuclear gene coding for this protein is therefore an important candidate for mutational detection studies in enzymatic complex I deficient patients. Screening of patient NDUFS2 cDNA by reverse transcriptase-polymerase chain reaction (RT-PCR) in combination with direct DNA sequencing revealed three missense mutations resulting in the substitution of conserved amino acids in three families.

Nuclear hnRNPA2B1 initiates and amplifies the innate immune response to DNA viruses

Abstract: INTRODUCTION Recognition of pathogen-derived nucleic acids by host cells is an evolutionarily conserved mechanism that induces immune defense responses to microbial infections. Most DNA viruses direct their genomic DNA into host cell nuclei, which can serve as an important molecular signature of DNA virus infection. However, little is known about the nuclear surveillance mechanisms for viral nucleic acids. RATIONALE Virus-induced type I interferon (IFN-I) expression depends on the TANK-binding kinase 1–interferon regulatory factor 3 (TBK1–IRF3) activation. We reasoned that nuclear DNA sensors may translocate to the cytoplasm to activate the TBK1–IRF3 pathway after recognizing viral DNA in the nucleus. Thus, we screened nuclear proteins that bound viral DNA and translocated from the nucleus to the cytoplasm after viral infection. Heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1) was identified as a potential DNA sensor. We then conducted a series of in vivo and in vitro experiments to probe the biological importance and activation mechanisms of hnRNPA2B1. Additionally, we explored its relationship with known cytosolic stimulator of interferon genes (STING)–dependent DNA sensors such as cyclic GAMP synthase (cGAS). RESULTS hnRNPA2B1 was found to bind viral DNA in the cell nucleus during herpes simplex virus–1 (HSV-1) infection. It then translocated to the cytoplasm and activated TBK1 through the tyrosine kinase Src. Accordingly, hnRNPA2B1 knockdowns and deficiency resulted in impaired DNA virus– but not RNA virus–induced IFN-I production and prolonged viral replication. The production of proinflammatory cytokines such as tumor necrosis factor–α (TNF-α) and interleukin-6 (IL-6) was unaffected. hnRNPA2B1 became dimerized after HSV-1 infection. Mutation of the dimer interface abrogated its nucleocytoplasmic translocation upon HSV-1 infection. Thus, hnRNPA2B1 dimerization is required for its nucleocytoplasmic translocation. Additionally, hnRNPA2B1 was demethylated at Arg226 after HSV-1 infection, which led to its activation and the subsequent initiation of IFN-β expression. This demethylation was catalyzed by the arginine demethylase JMJD6. hnRNPA2B1 with dimer interface mutation was unable to associate with JMJD6 after HSV-1 infection and showed increased amounts of arginine methylation compared to full-length hnRNPA2B1, indicating that dimerization was required for its demethylation. To probe the relationship between hnRNPA2B1 and the recognized DNA sensor pathways, we found that the overexpression of hnRNPA2B1 increased HSV-1–induced TBK1 activation and Ifnb1 expression in Cgas–/– L929 cells. Thus, hnRNPA2B1 could induce IFN-I in a cGAS-independent manner at least in part. This is consistent with earlier evidence suggesting the existence of other IFN-I–initiating molecules in the innate response against DNA virus. Wild-type macrophages showed higher and more sustained Ifnb1 expression than Hnrnpa2b1–/– macrophages in response to DNA viruses. Thus, hnRNPA2B1 was required for fully activating type I interferon production against DNA viruses mediated by cGAS, interferon-γ–inducible protein 16 (IFI16), and STING pathways. Mechanistically, hnRNPA2B1 bound CGAS, IFI16,and STING mRNAs and promoted their nucleocytoplasmic trafficking to amplify cytoplasmic innate sensor signaling. The translation of these mRNAs was impaired in the absence of hnRNPA2B1 after HSV-1 infection. hnRNPA2B1 was constitutively associated with fat mass and obesity-associated protein (FTO). This association was abrogated after HSV-1 infection. By this means, hnRNPA2B1 promoted the N6-methyladenosine (m6A) modification and nucleocytoplasmic trafficking of CGAS, IFI16,and STING mRNAs. Thus, hnRNPA2B1 facilitates the efficient induction of antiviral IFN-I production mediated by cGAS, IFI16, and STING. CONCLUSION We identified hnRNPA2B1 as an innate sensor that initiates type I IFN production upon DNA virus infection in the nucleus. hnRNPA2B1 also amplifies type I IFN responses by directly enhancing STING-dependent cytosolic DNA sensing pathways.