Effect of histones from brain on DNA-synthesis in vitro

Summary The results of these experiments demonstrate that histones from brain inhibit the replication of DNA in vitro. A similar effect is observed with polylysine or polyarginine. The reversion of inhibition by polyglutamic acid or acidic proteins is completed in all cases except when the DNA is previously complexed with histones, polyarginine or polylysine. This suggest that histones masking of DNA towards the polymerases involves electrostatic forces.These studies were supported by the Consejo Nacional de Investigaciones Científicas y Técnicas and the Instituto Nacional de Farmacología y Bromatología, Argentina.Abbreviations. DNA-polymerase: deoxynucleoside triphosphate: DNA deoxynucleotidyl transferase (E.C. 2.7.7.7.).     

Inhibitory effect of exogenous histones on cellular growth and DNA synthesis in Chlamydomonas reinhardtii.]

The addition of exogenous histones to synchronous culture of Chlamydomonas reinhardtii, at the beginning of the cell cycle, leads to the death of the cells. The same amount of histones added in the middle of the cycle, only blocks the cell division. The mechanism of this inhibition effect of the histones could involve a block at the level of the chloroplast DNA replication.     

Chromatin assembly during S phase: contributions from histone deposition, DNA replication and the cell division cycle

During S phase of the eukaryotic cell division cycle, newly replicated DNA is rapidly assembled into chromatin. Newly synthesised histones form complexes with chromatin assembly factors, mediating their deposition onto nascent DNA and their assembly into nucleosomes. Chromatin assembly factor 1, CAF-1, is a specialised assembly factor that targets these histones to replicating DNA by association with the replication fork associated protein, proliferating cell nuclear antigen, PCNA. Nucleosomes are further organised into ordered arrays along the DNA by the activity of ATP-dependent chromatin assembly and spacing factors such as ATP-utilising chromatin assembly and remodelling factor ACF. An additional level of controlling chromatin assembly pathways has become apparent by the observation of functional requirements for cyclin-dependent protein kinases, casein kinase II and protein phosphatases. In this review, we will discuss replication-associated histone deposition and nucleosome assembly pathways, and we will focus in particular on how nucleosome assembly is linked to DNA replication and how it may be regulated by the cell cycle control machinery.     

Microtubule associated protein tau binds to double-stranded but not single-stranded DNA

Tau, a major microtubule-associated protein of the neuron, which is known to promote the assembly of and to stabilize microtubules, has also been seen associated with chromatin in neuronal cell lines, but its role in this subcellular compartment is still unknown. In this study, the binding of tau to DNA was investigated using the electrophoretic mobility shift assay. Using polynucleotide as probe, we found that tau bound to double-stranded but not to single-stranded DNA. Formation of tau-polynucleotide complex was disrupted by alkaline pH and a high concentration of NaCl, but was not affected by dithiothreitol. Electron microscopy revealed that the protein associated with the nucleic acid in a necklacelike manner. DNA-cellulose chromatography and radioimmunodot-blot analyses showed that calf thymus histones VI-S, VII-S and VIII-S could replace both recombinant human brain tau₃₅₂ (tau-23) and tau₄₄₁ (tau-40) from DNA. Thus, tau appears to bind to DNA reversibly in the presence of histones. Received 24 November 2002; received after revision 28 December 2002; accepted 30 December 2002 RID="*" ID="*"Corresponding author.     

Histone hydrolase in tadpole liver

Résumé Le cytoplasme du foie de têtard contient une enzyme qui hydrolyse spécifiquement les protéines basiques telle que la polylysine, la protamine et les histones. L'addition d'autres protéines telles que l'albumine, la globuline et l'acide polyglutamique inhibe l'activité de cette enzyme. Les résultats obtenus montrent que cette hydrolysase extraite du foie de têtard diffère des cathépsines ou de l'histone-hydrolase du rein de rat.

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This work was supported by research grants from the National Institute of Arthritis and Metabolic Diseases No. AM 09602, National Science Foundation No. GB-22663 and from the National Cancer Institute No. CA 07174.     

Implication of the VRK1 chromatin kinase in the signaling responses to DNA damage: a therapeutic target?

DNA damage causes a local distortion of chromatin that triggers the sequential processes that participate in specific DNA repair mechanisms. This initiation of the repair response requires the involvement of a protein whose activity can be regulated by histones. Kinases are candidates to regulate and coordinate the connection between a locally altered chromatin and the response initiating signals that lead to identification of the type of lesion and the sequential steps required in specific DNA damage responses (DDR). This initiating kinase must be located in chromatin, and be activated independently of the type of DNA damage. We review the contribution of the Ser-Thr vaccinia-related kinase 1 (VRK1) chromatin kinase as a new player in the signaling of DNA damage responses, at chromatin and cellular levels, and its potential as a new therapeutic target in oncology. VRK1 is involved in the regulation of histone modifications, such as histone phosphorylation and acetylation, and in the formation of γH2AX, NBS1 and 53BP1 foci induced in DDR. Induction of DNA damage by chemotherapy or radiation is a mainstay of cancer treatment. Therefore, novel treatments can be targeted to proteins implicated in the regulation of DDR, rather than by directly causing DNA damage.     

Biotin as a regulator of some haematic parameters and of DNA-content of the liver of old rats

Biotin administration to old rats (28 months) causes in the blood an increase of ATP, glucose, triglycerides, alkaline phosphatase and a decrease of cholesterol and acid phosphatase; in the liver DNA and electrostatic interactions between DNA and histones are increased. Such parameters come within the values shown by adult rats.     

Biotin as a regulator of some haematic parameters and of DNA-content of the liver of old rats

Summary Biotin administration to old rats (28 months) causes in the blood an increase of ATP, glucose, triglycerides, alkaline phosphatase and a decrease of cholesterol and acid phosphatase; in the liver DNA and electrostatic interactions between DNA and histones are increased. Such parameters come within the values shown by adult rats.     

Bridging epigenomics and complex disease: the basics

The DNA sequence largely defines gene expression and phenotype. However, it is becoming increasingly clear that an additional chromatin-based regulatory network imparts both stability and plasticity to genome output, modifying phenotype independently of the genetic blueprint. Indeed, alterations in this “epigenetic” control layer underlie, at least in part, the reason for monozygotic twins being discordant for disease. Functionally, this regulatory layer comprises post-translational modifications of DNA and histones, as well as small and large noncoding RNAs. Together these regulate gene expression by changing chromatin organization and DNA accessibility. Successive technological advances over the past decade have enabled researchers to map the chromatin state with increasing accuracy and comprehensiveness, catapulting genetic research into a genome-wide era. Here, aiming particularly at the genomics/epigenomics newcomer, we review the epigenetic basis that has helped drive the technological shift and how this progress is shaping our understanding of complex disease.     

Chromatin circles in amphibian previtellogenic oocytes

Summary Previtellogenic oocytes ofOdontophrynus americanus display hundreds of chromatin circles. Electron microscopy of spread preparations of isolated nuclei shows that the circles originate from the chromatin. The circles change their morphology and form new copies. The length of the DNA packed in the nucleosomal circles is about 2.5–3.5 m or multiples of this value. Assuming that histones need not be removed from chromatin before DNA replication³ we suggest that the circles might belong to the process of rDNA amplification.This work was supported by grants from the Brazilian CNPq, FAPESP and FEDIB.We thank Dr A. Brunner Jr for the use of the electron microscope and Dr N. Leon for his valuable help.     

Internalization of cationic peptides: the road less (or more?) traveled

Facilitating the entry of molecules into mammalian cells is of great interest to fields as diverse as cell biology and drug delivery. The discovery of natural protein transduction domains and the development of artificial ones, including polyarginine, provides a means to achieve this goal. Here, we comment on key chemical and biological aspects of cationic peptide internalization, including the physiological relevance of this process. Received 13 April 2006; received after revision 21 May 2006; accepted 2 June 2006