FoxA2 and RNA Pol II Mediate Human Islet Amyloid Polypeptide Turnover in ER-stressed Pancreatic β-cells

Here, we investigated transcriptional and trafficking mechanisms of human islet amyloid polypeptide (hIAPP) in normal and stressed β-cells. In high glucose-challenged human islets and rat insulinoma cells overexpressing hIAPP, cell fractionation studies revealed increased accumulation of hIAPP. Unexpectedly, a significant fraction (up to 22%) of hIAPP was found in the nuclear soluble and chromatin-enriched fractions of cultured human islet and rat insulinoma cells. The nucleolar accumulation of monomeric forms of hIAPP did not have any adverse effect on the proliferation of β-cells nor did it affect nucleolar organization or function. However, intact nucleolar organization and function were essential for hIAPP expression under normal and ER-stress conditions as RNA polymerase II inhibitor, α-amanitin, reduced hIAPP protein expression evoked by high glucose and thapsigargin. Promoter activity studies revealed the essential role of transcription factor FoxA2 in hIAPP promoter activation in ER-stressed β-cells. Transcriptome and secretory studies demonstrate that the biosynthetic and secretory capacity of islet β-cells was preserved during ER stress. Thus, the main reason for increased intracellular hIAPP accumulation is its enhanced biosynthesis under these adverse conditions.

Nucleophosmin in Its Interaction with Ligands

Nucleophosmin (NPM1) is a mainly nucleolar protein that shuttles between nucleoli, nucleoplasm and cytoplasm to fulfill its many functions. It is a chaperone of both nucleic acids and proteins and plays a role in cell cycle control, centrosome duplication, ribosome maturation and export, as well as the cellular response to a variety of stress stimuli. NPM1 is a hub protein in nucleoli where it contributes to nucleolar organization through heterotypic and homotypic interactions. Furthermore, several alterations, including overexpression, chromosomal translocations and mutations are present in solid and hematological cancers. Recently, novel germline mutations that cause dyskeratosis congenita have also been described. This review focuses on NPM1 interactions and inhibition. Indeed, the list of NPM1 binding partners is ever-growing and, in recent years, many studies contributed to clarifying the structural basis for NPM1 recognition of both nucleic acids and several proteins. Intriguingly, a number of natural and synthetic ligands that interfere with NPM1 interactions have also been reported. The possible role of NPM1 inhibitors in the treatment of multiple cancers and other pathologies is emerging as a new therapeutic strategy.

A Fob1 independent role for the Smc5/6 complex in the rDNA that modulates lifespan

SUMMARYThe ribosomal DNA (rDNA) in Saccharomyces cerevisiae is in one tandem repeat array on Chromosome XII. Two spacer regions within each repetitive element, called non-transcribed spacer 1 (NTS1) and NTS2, are important in nucleolar organization. Smc5/6 localizes to both NTS1 and NTS2 and is involved in the regulation of Sir2 and Cohibin binding at NTS1, whereas Fob1 and Sir2 are required for optimal binding of the complex to NTS1 and NTS2, respectively. We demonstrate that Smc5/6 functions in chromatin silencing at NTS1 independently of its role in homologous recombination (HR) when forks pause at the replication fork barrier (RFB). In contrast, when the complex does not localize to the rDNA in nse3-1 mutants, the shortened lifespan correlates with NTS2 homeostasis independently of FOB1 status. Our data identify the importance of Smc5/6 integrity in NTS2 transcriptional silencing and repeat tethering, which in turn underscores rDNA stability and replicative lifespan.HighlightsSmc5/6 is important for transcriptional silencing in the rDNA.Smc5/6 tethers the rDNA array to the periphery.Transcriptional silencing of ncRNA at NTS1 and NTS2 is differentially regulated.Smc5/6 has a role in rDNA maintenance independent of HR processing at the RFB.Fob1-independent disruption of Smc5/6 at NTS2 leads to lifespan reduction.

Presence of 15p Marker D15Z1 on the Short Arm of Acrocentric Chromosomes is Associated with Aneuploid Offspring in Mexican Couples

Variation in the location of the 15p region D15Z1 is recognized as a polymorphism in several human populations. We used high-stringency Fluorescence In Situ Hybridization (FISH) to detect D15Z1 in a Mexican cohort. Here, we report the presence of extra D15Z1 sequences on the p-arm of acrocentric chromosomes other than 15 in two groups of Mexican couples, one with healthy offspring (n = 75) and the other with aneuploid offspring (n = 87), mainly trisomy 21. The additional D15Z1 polymorphism was significantly increased in individuals with aneuploid offspring (26.4%), in comparison to individuals with healthy offspring (14%). The most frequent acceptor chromosome of D15Z1 was chromosome 13p, followed by 14p, and finally, 21p. Our results show an overall frequency of 21.6% of this polymorphism in the Mexican population and suggest that its presence might be associated with the mis-segregation of other acrocentric chromosomes and aneuploid offspring. The high frequency of the polymorphism of the D15Z1 sequence on acrocentric chromosomes other than 15 suggests a sequence homogenization of the acrocentric p arms, related to the important function of the centromere and the nucleolar organization region, which flank satellite III DNA.

Localización de satélites y cromosomas NOR para la interpretación del cariotipo de Sesbania virgata (Papilionoideae, Sesbanieae) de dos poblaciones americanas

<p><strong>Background</strong>: Cytogenetic studies in the genus <em>Sesbania</em> show lack of agreement among the researchers about the precise number and position of secondary constrictions and satellites as well as their relation to the organization of the nucleolus. The lack of this information makes it difficult to carry out reliable comparative cytogenetic studies and chromosome evolution in this genus.</p><p><strong>Questions</strong>: Where are the secondary constrictions and satellites located in the chromosomes of <em>Sesbania</em> <em>virgat</em>a? Do these regions actively participate in the nucleolar organization?</p><p><strong>Study species</strong>: <em>Sesbania virgate</em> (Cav.) Pers.</p><p><strong>Study site</strong>: Municipality of Tlacotalpan, Mexico and Province of Salta, Argentina.</p><p><strong>Methods</strong>: Surface spreading and air-drying technique was applied to obtain chromosomes in prometaphase and typical metaphase from radicular meristems.</p><p><strong>Results</strong>: Each population exhibited a different karyotype and only two secondary constrictions associated with microsatellites in the short arms of the smallest chromosome pair and not in long arms as was suggested by other authors. The inclusion of secondary constrictions and satellites in the nucleolus of cells in prometaphase allowed to corroborate their active participation in the formation of this one. This information was used to reevaluate the position of the nucleolar organizer regions "NOR´s".</p><strong>Conclusions</strong>: Our results agree with the predominant point of view on the location of the "NOR´s" in the short arms of plant species, particularly in legumes. In addition, given that the populations under study are geographically isolated, we suggest that an active process of speciation manifested in the two found cytotypes whose differences are attributed to changes in the proportion of arms of the satellite chromosomes is favored.