SETDB1 regulates microtubule dynamics

ObjectivesSETDB1 is a methyltransferase responsible for the methylation of histone H3‐lysine‐9, which is mainly related to heterochromatin formation. SETDB1 is overexpressed in various cancer types and is associated with an aggressive phenotype. In agreement with its activity, it mainly exhibits a nuclear localization; however, in several cell types a cytoplasmic localization was reported. Here we looked for cytoplasmic functions of SETDB1.MethodsSETDB1 association with microtubules was detected by immunofluorescence and co‐sedimentation. Microtubule dynamics were analysed during recovery from nocodazole treatment and by tracking microtubule plus‐ends in live cells. Live cell imaging was used to study mitotic kinetics and protein–protein interaction was identified by co‐immunoprecipitation.ResultsSETDB1 co‐sedimented with microtubules and partially colocalized with microtubules. SETDB1 partial silencing led to faster polymerization and reduced rate of catastrophe events of microtubules in parallel to reduced proliferation rate and slower mitotic kinetics. Interestingly, over‐expression of either wild‐type or catalytic dead SETDB1 altered microtubule polymerization rate to the same extent, suggesting that SETDB1 may affect microtubule dynamics by a methylation‐independent mechanism. Moreover, SETDB1 co‐immunoprecipitated with HDAC6 and tubulin acetylation levels were increased upon silencing of SETDB1.ConclusionsTaken together, our study suggests a model in which SETDB1 affects microtubule dynamics by interacting with both microtubules and HDAC6 to enhance tubulin deacetylation. Overall, our results suggest a novel cytoplasmic role for SETDB1 in the regulation of microtubule dynamics.

SETDB1 association with microtubules inhibits microtubule polymerization and enhances their instability. SETDB1 may affect the microtubules by interacting with HDAC6 to enhance HDAC6 tubulin deacetylation activity.     

Accelerated Biodegradation of Cement by Sulfur-Oxidizing Bacteria as a Bioassay for Evaluating Immobilization of Low-Level Radioactive Waste

Disposal of low-level radioactive waste by immobilization in cement is being evaluated worldwide. The stability of cement in the environment may be impaired by sulfur-oxidizing bacteria that corrode the cement by producing sulfuric acid. Since this process is so slow that it is not possible to perform studies of the degradation kinetics and to test cement mixtures with increased durability, procedures that accelerate the biodegradation are required. Semicontinuous cultures of Halothiobacillus neapolitanus and Thiomonas intermedia containing thiosulfate as the sole energy source were employed to accelerate the biodegradation of cement samples. This resulted in a weight loss of up to 16% after 39 days, compared with a weight loss of 0.8% in noninoculated controls. Scanning electron microscopy of the degraded cement samples revealed deep cracks, which could be associated with the formation of low-density corrosion products in the interior of the cement. Accelerated biodegradation was also evident from the leaching rates of Ca²⁺ and Si²⁺, the major constituents of the cement matrix, and Ca exhibited the highest rate (up to 20 times greater than the control rate) due to the reaction between free lime and the biogenic sulfuric acid. Leaching of Sr²⁺ and Cs⁺, which were added to the cement to simulate immobilization of the corresponding radioisotopes, was also monitored. In contrast to the linear leaching kinetics of calcium, silicon, and strontium, the leaching pattern of cesium produced a saturation curve similar to the control curve. Presumably, the leaching of cesium is governed by the diffusion process, whereas the leaching kinetics of the other three ions seems to governed by dissolution of the cement.     

Akustische Barrieren beim Waldbauml?ufer (Certhia familiaris)?

Zusammenfassung Waldbaumläufer besitzen je nach geographischer Herkunft deutlich verschiedene Reviergesangs-Strophen. Zusammenfassen lassen sich jene aus Mittel- und W-Europa (Certhia familiaris macrodactyla, mWb;C. f. britannica undC. f. corsa), aus dem weiteren Himalaya-Gebiet (Nepal,C. f. mandellii, nWb; und SW-China,C. f. khamensis, cwb) und als weitere Gruppe die aus Japan (C. f. orientalis, jWb). Differenzen im Gesang bestehen hinsichtlich der Zahl der Elemente in der Strophe, Frequenzumfang der Strophe, Frequenzverlauf der Elemente und deren Frequenzumfang. Der nWb reagiert nicht auf Gesang des mWb, umgekehrt besteht hohe Reaktionsfreudigkeit des mWb auf Str. des nWb. Freiland-Experimente mit unveränderten, gekürzten und künstlichen Str. aus gereihten Einzel-El. zeigen, daß bestimmter Frequenz-Verlauf der El. die Reaktion hervorruft. Es sind solche El. des nWb, die beim mWb in ähnlicher Form auftreten. Vielfach sind es aber El., die zwar vordergründig geringe oder keine Übereinstimmungen aufweisen, in einzelnen El.-Abschnitten jedoch erkennbare Frequenz-Entsprechungen besitzen. Einzelne dieser El. von mWb und nWb sind als homolog zu betrachten. In den meisten Fällen kann über die mögliche Homologie nicht entschieden werden, denn über El.-Veränderungen in der Evolution akustischer Signale ist bei Baumläufern zu wenig bekannt.

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Acoustic barriers in the Tree Creeper (Certhia familiaris)?

Summary Tree Creepers use according to their distributional origin different territorial songs. Populations with similar songs occur in Central and W Europe (C. f. macrodactyla, mWb;C. f. britannica, C. f. corsa), in Himalayan East Asia (Nepal,C. f. mandellii, nWb; SW China,C. f. khamensis, cWb) and Japan (C. f. orientalis, jWb). Differences in the territorial songs refer to number of elements in the verse, frequency volume of the verse, frequency modulation of the elements and their frequency volume. The Tree Creeper from Nepal (mandellii) does not react on the song of Central European Tree Creepers (macrodactyla), but the latter is very responsive tomandellii verses. Field experiments using unchanged, shortened or artificial verses consisting of one natural element arranged in a row, demonstrate that only certain frequency modulations evoke reactions. Such elements occur at least in similar expression in nWb and mWb as well. Often also such elements are answered which at first sight do not correspond or only to a low degree to mWb elements, but do so in certain small sections of mWb elements. Several of the similar elements in mWb and nWb are to be considered homologous. But in most cases where reaction in field experiments is high, the homology of the elements concerned cannot be substantiated. Changes of element structures in the course of vocal evolution in the [familiaris] superspecies is too poorly known.

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Results of the Himalaya Expeditions ofJ. Martens, No. 146. — For No. 145 see: Stuttgarter Beitr. Naturk., (A) 411: 1–43, 1987. — J. M. sponsored by Deutscher Akademischer Austauschdienst and Deutsche Forschungsgemeinschaft.     

A mutation in aminopeptidase N (CD13) isolated from a patient suffering from leukemia leads to an arrest in the endoplasmic reticulum

Human aminopeptidase N (APN) is used as a routine marker for myelomonocytic cells in hematopoietic malignant disorders. Its gene and surface expressions are increased in cases of malignant transformation, inflammation, or T cell activation, whereas normal B and resting T cells lack detectable APN protein expression. In this study we elucidated the intracellular distribution, expression pattern, and enzymatic activity of a naturally occurring mutation in the coding region of the APN gene. At physiological temperatures the mutant protein is enzymatically inactive, persists as a mannose-rich polypeptide in the endoplasmic reticulum, and is ultimately degraded by an endoplasmic reticulum-associated degradation pathway. It shows in part the distinct behavior of a temperature-sensitive mutant with a permissive temperature of 32 degrees C, leading to correct sorting of the Golgi compartment accompanied by the acquisition of proper glycosylation but without reaching the cell-surface membrane and without regaining its enzymatic activity. Because the patient bearing this mutation suffered from leukemia, possible links to the pathogenesis of leukemia are discussed.     

Transgenerational maintenance of transgene body CG but not CHG and CHH methylation

In plants, RNA-directed DNA methylation (RdDM) can target both transgene promoters and coding regions/gene bodies. RdDM leads to methylation of cytosines in all sequence contexts: CG, CHG and CHH. Upon segregation of the RdDM trigger, at least CG methylation can be maintained at promoter regions in the progeny. So far, it is not clear whether coding region methylation can be also maintained. We showed that the body of Potato spindle tuber viroid (PSTVd) transgene constructs became densely de novo methylated at CG, CHG and CHH sites upon PSTVd infection. In this study, we demonstrate that in viroid-free progeny plants, asymmetric CHH and CHG methylation was completely lost. However, symmetric CG methylation was stably maintained for at least two generations. Importantly, the presence of transgene body methylation did not lead to an increase of dimethylation of histone H3 lysine 9 or a decrease of acetylation of H3. Our data supports the view that CG methylation can be maintained not only in promoters but also in the body of transgenes. They further suggest that maintenance of methylation may occur independently of tested chromatin modifications.