The STING pathway and regulation of innate immune signaling in response to DNA pathogens

The innate immune system has evolved a variety of sensing mechanisms to detect and counter microbial invasion. These include the Toll-like receptor (TLR), cytoplasmic, nucleotide binding oligomerization domain (NOD)-like receptor and RIG-I-like helicase (RLH) pathways. However, how the cell detects pathogen-associated DNA to trigger host defense, including the production of interferon, remains to be fully clarified. Understanding these processes could have profound implications into how we understand and treat a variety of microbial-related disease, including viral-associated cancers, as well as autoimmune disorders. Recently, an endoplasmic reticulum-associated molecule referred to as STING (for stimulator of interferon genes) was isolated and shown to be critical for regulating the production of IFN in response to cytoplasmic DNA. Here, we review recent discoveries relating to the detection of foreign DNA, including the importance of the STING and inflammasome pathways and the triggering of innate signaling processes.     

Esculentin(1-21), an amphibian skin membrane-active peptide with potent activity on both planktonic and biofilm cells of the bacterial pathogen Pseudomonas aeruginosa

Pseudomonas aeruginosa is an opportunistic bacterial pathogen that forms sessile communities, named biofilms. The non-motile forms are very difficult to eradicate and are often associated with the establishment of persistent infections, especially in patients with cystic fibrosis. The resistance of P. aeruginosa to conventional antibiotics has become a growing health concern worldwide and has prompted the search for new anti-infective agents with new modes of action. Naturally occurring antimicrobial peptides (AMPs) represent promising future template candidates. Here we report on the potent activity and membrane-perturbing effects of the amphibian AMP esculentin(1-21), on both the free-living and sessile forms of P. aeruginosa, as a possible mechanism for biofilm disruption. Furthermore, the findings that esculentin(1-21) is able to prolong survival of animals in models of sepsis and pulmonary infection indicate that this peptide can be a promising template for the generation of new antibiotic formulations to advance care of infections caused by P. aeruginosa.     

The BLM dissolvasome in DNA replication and repair

RecQ DNA helicases are critical for proper maintenance of genomic stability, and mutations in multiple human RecQ genes are linked with genetic disorders characterized by a predisposition to cancer. RecQ proteins are conserved from prokaryotes to humans and in all cases form higher-order complexes with other proteins to efficiently execute their cellular functions. The focus of this review is a conserved complex that is formed between RecQ helicases and type-I topoisomerases. In humans, this complex is referred to as the BLM dissolvasome or BTR complex, and is comprised of the RecQ helicase BLM, topoisomerase IIIα, and the RMI proteins. The BLM dissolvasome functions to resolve linked DNA intermediates without exchange of genetic material, which is critical in somatic cells. We will review the history of this complex and highlight its roles in DNA replication, recombination, and repair. Additionally, we will review recently established interactions between BLM dissolvasome and a second set of genome maintenance factors (the Fanconi anemia proteins) that appear to allow coordinated genome maintenance efforts between the two systems.     

Sea urchin embryo as a model for analysis of the signaling pathways linking DNA damage checkpoint,DNA repair and apoptosis

DNA integrity checkpoint control was studied in the sea urchin early embryo. Treatment of the embryos with genotoxic agents such as methyl methanesulfonate (MMS) or bleomycin induced the activation of a cell cycle checkpoint as evidenced by the occurrence of a delay or an arrest in the division of the embryos and an inhibition of CDK1/cyclin B activating dephosphorylation. The genotoxic treatment was shown to induce DNA damage that depended on the genotoxic concentration and was correlated with the observed cell cycle delay. At low genotoxic concentrations, embryos were able to repair the DNA damage and recover from checkpoint arrest, whereas at high doses they underwent morphological and biochemical changes characteristic of apoptosis. Finally, extracts prepared from embryos were found to be capable of supporting DNA repair in vitro upon incubation with oligonucleotides mimicking damage. Taken together, our results demonstrate that sea urchin early embryos contain fully functional and activatable DNA damage checkpoints. Sea urchin embryos are discussed as a promising model to study the signaling pathways of cell cycle checkpoint, DNA repair and apoptosis, which upon deregulation play a significant role in the origin of cancer. Received 10 April 2007; accepted 23 April 2007     

CD20-related signaling pathway is differently activated in normal and dystrophic circulating CD133+ stem cells

Among the heterogeneous population of circulating hematopoietic and endothelial progenitors, we identified a subpopulation of CD133⁺ cells displaying myogenic properties. Unexpectedly, we observed the expression of the B-cell marker CD20 in blood-derived CD133⁺ stem cells. The CD20 antigen plays a role in the modulation of intracellular calcium homeostasis through signaling pathways activation. Several observations suggest that an increase in intracellular calcium concentration ([Ca²⁺]_i) could be involved in the etiology of the Duchenne muscular dystrophy (DMD). Here, we show that a CD20-related signaling pathway able to induce an increase in [Ca²⁺]_i is differently activated after brain derived neurotrophic factor (BDNF) stimulation of normal and dystrophic blood-derived CD133⁺ stem cells, supporting the assumption of a “CD20-related calcium impairment-affecting dystrophic cells. Presented findings represent the starting point toward the expansion of knowledge on pathways involved in the pathology of DMD and in the behavior of dystrophic blood-derived CD133⁺ stem cells. Received 15 October 2008; received after revision 27 November 2008; accepted 05 December 2008     

The changes of the pentose phosphate pathway in red cells during their ageingin vitro

Zusammenfassung Imin vitro alternden Blut wird Abnahme der gesamten Pentosemenge sowie Abnahme deren Bildung während der Inkubation mit Glucose fest-gestellt. Das Absinken ist grösser als die gleichzeitige Abnahme der Glucoseauswertung. Ebenfalls nimmt die stimulatorische Wirkung des Methylenblaus auf die Pentosebildung während der Alterung des Blutes stark ab.     

The mode of timing of DNA replication and of mitosis in cultured animal cells

Zusammenfassung Eine indirekte Methode der Analyse des mitotischen Zell-Zyklus wurde an verschieden rasch, jedoch stets exponentiell wachsenden Suspensions-Kulturen von neoplastischen Maus-Mast-Zellen des Stammes P815Y und Kulturen von L929 Maus-Zellen geprüft. Die experimentellen Kurven (Dauer derG2-periode als Funktion der Kulturen-Verdopplungszeit, mitotischer Index als Funktion der spezifischen Wachstumsgeschwindigkeit der Kulturen, und DNA-synthetischer Index als Funktion der spezifischen Wachstumsgeschwindigkeit) sind mit der Hypothese im Einklang, nach der die Dauern derG1-,S-,G2- und derM-Periode homogen-lineare Funktionen der Generationsdauer respektive der Gesamtdauer des Zell-Zyklus sind. Demzufolge sind unter Bedingungen streng exponentiellen Kulturenwachstums die DNA-Synthese-Periode, dieG2-Periode und die Mitose-Periode nicht von konstanter Dauer, wie üblicherweise angenommen wird. Die experimentell erzwungene Verlängerung oder Verkürzung der Generationsdauer der sich in der Kultur teilenden Zellen (R-Zellen) wird somit nicht allein durch dieG1-Periode bestimmt, sondern durch gleichzeitige und prozentual gleichmässige Expansion oder Kontraktion aller vier charakteristischen Phasen des Zell-Zyklus.

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This investigation was supported by the Office of Naval Research under contract No. Nonr-266(76) and by the Health Research Council of the City of New York under contract No. I-428, and was carried out at Columbia University (Department of Biochemistry).

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Recipient of Career Scientist Award of the Health Research Council of the City of New York under contract No. I-428.

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This paper has been dedicated to my former teacher Prof.Ad. Portmann on the occasion of his 70th birthday.     

The effect of DDT on DNA replication in the larval salivary gland cells ofDrosophila melanogaster

Summary The inhibitory effect of DDT on the initial stage of the DNA replication process in polytene chromosomes of larval salivary gland cells ofDrosophila melanogaster was investigated and possible mechanisms for the inhibition are discussed.     

Proliferation of spleen cells from mice infected with friend virus in the spleens of unirradiated and irradiated mice

Résumé Quand des cellules spléniques de souris CBA/HT₆T₆ infectées par le virus de Friend sont injectées à des souris CBA/H normales, les rates des receveurs s'hypertrophient au cours de la semaine suivante. Une analyse chromosomale montra que toutes les cellules en division provenaient de l'hôte, suggérant que la splénomégalie fait suite à la libération du virus des cellules injectées.     

Migration of lymphoid cells to the bone marrow of rat following eradication of cells in DNA synthesis and in mitosis

Summary Eradication of replicating bone marrow cells of rat by means of combined administration of single doses of hydroxyurea and vinblastin is followed within 9–10 h by an inflow of lymphoid cells of extramedullary origin in the range of 13,200,000/femur. The rat bone marrow with a high content of lymphoid cells was previously shown to be concentrated in stem cells. The factor(s) which convey the information of decrease of replicating marrow cells to extramedullary sites is at present unknown.Acknowledgment. This study was supported by a grant from the Chief Scientist Office, Ministry of Health, Israel.Hydroxyurea for this investigation was given as a gift by the Squibb Institute of Medical Research, Princeton, N.J. USA, to which the authors are indebted.