Cohesin is a protein complex known for its essential role in chromosome segregation. However, cohesin and associated factors have additional functions in transcription, DNA damage repair, and chromosome condensation. The human cohesinopathy diseases are thought to stem not from defects in chromosome segregation but from gene expression. The role of cohesin in gene expression is not well understood. We used budding yeast strains bearing mutations analogous to the human cohesinopathy disease alleles under control of their native promoter to study gene expression. These mutations do not significantly affect chromosome segregation. Transcriptional profiling reveals that many targets of the transcriptional activator Gcn4 are induced in the eco1-W216G mutant background. The upregulation of Gcn4 was observed in many cohesin mutants, and this observation suggested protein translation was reduced. We demonstrate that the cohesinopathy mutations eco1-W216G and smc1-Q843 Δ are associated with defects in ribosome biogenesis and a reduction in the actively translating fraction of ribosomes, eiF2α-phosphorylation, and 35 S-methionine incorporation, all of which indicate a deficit in protein translation. Metabolic labeling shows that the eco1-W216G and smc1-Q843 Δ mutants produce less ribosomal RNA, which is expected to constrain ribosome biogenesis. Further analysis shows that the production of rRNA from an individual repeat is reduced while copy number remains unchanged. Similar defects in rRNA production and protein translation are observed in a human Roberts syndrome cell line. In addition, cohesion is defective specifically at the rDNA locus in the eco1-W216G mutant, as has been previously reported for Roberts syndrome. Collectively, our data suggest that cohesin proteins normally facilitate production of ribosomal RNA and protein translation, and this is one way they can influence gene expression. Reduced translational capacity could contribute to the human cohesinopathies.

Chromosome cohesion is a cell-cycle-regulated process in which sister chromatids are held together from the time of replication until the time of separation at the metaphase-to-anaphase transition, ensuring accurate chromosome segregation [1-9]. Chromosome cohesion is established during S phase, and this process requires the four subunits of the cohesin complex (Smc1, Smc3, Mcd1/Scc1, and Irr1/Scc3) and the acetyltransferase Eco1 [10-13]. Acetylation of Smc3 by Eco1 at two evolutionarily conserved lysine residues promotes cohesion establishment during S phase in budding yeast and humans [14-16]. Here we report that Hos1, a member of the evolutionarily conserved class I histone deacetylase family, acts as a deacetylase for Smc3 in S. cerevisiae. We examine the Smc3 acetylation level in nine histone deacetylase deletion strains and find that the acetylation level is increased specifically in a hos1Δ strain post-S phase. Coimmunoprecipitation experiments show that Hos1 interacts with Smc3 and that the interaction is most pronounced as cells reach anaphase. We provide direct evidence that Hos1 can deacetylate Smc3 and retains a soluble pool of deacetylated Smc3. Overexpression of Hos1 results in less acetylation of Smc3 and cohesion defects in both WT and eco1 mutant strains; mutation of the Hos1 active site abolishes the defects. Hos1 may help to maintain a pool of unacetylated Smc3 that can be used for new chromosome cohesion.

The cohesin network has an essential role in chromosome segregation, but also plays a role in DNA damage repair. Eco1 is an acetyltransferase that targets subunits of the cohesin complex and is involved in both the chromosome segregation and DNA damage repair roles of the network. Using budding yeast as a model system, we find that mutations in Eco1, including a genocopy of a human Roberts syndrome allele, do not cause gross defects in chromosome cohesion. We examined how mitotic and meiotic DNA damage repair is affected by mutations in Eco1. Strains containing mutations in Eco1 are sensitive to DNA damaging agents that cause double-strand breaks, such as X-rays and bleomycin. While meiotic crossing over is relatively unaffected in strains containing the Roberts mutation, reciprocal mitotic crossovers occur with extremely low frequency in this mutant background. Our results suggest that Eco1 promotes the reciprocal exchange of chromosome arms and maintenance of heterozygosity during mitosis.

Mutations in the cohesin acetyltransferase Eco1 or the cohesin ring compromise nucleolar function in budding yeast. A mutation in Eco1 that is associated with the human disease Roberts syndrome compromises looping interactions at the ribosomal DNA and transcription. Depletion of cohesion in a single cell cycle disrupts nucleolar integrity.

The geomorphic expression of the frontal Western Foothills in central Taiwan is usually defined by a late Holocene scarp that ranges from tens to hundreds of meters in height. This scarp is the product of displacement on a near-surface 20-30Њ east-dipping thrust fault, the Chelungpu fault, which ruptured during the Chichi earthquake. The large scarp height may correspond directly to the accumulation of successive surface ruptures. The Chushan site is located on the southern part of this fault where the Chichi earthquake produced a scarp with a 1.7-m vertical offset for the total vertical separation. Based on core-boring estimates, the vertical displacement on both sides is 7 m along a 24Њ east-dipping thrust fault. The results from our paleoseismic analysis indicate that five large paleoearthquake events have caused the large offsets during the past 2 ka. The radiocarbon age constraints of the paleoearthquakes suggest a clustering of 540-790 cal yr B.P. (E2), 710-950 cal yr B.P. (E3), 1380-1700 cal yr B.P. (E4), 1710-1930 cal yr B.P. (E5), and the 1999 Chichi earthquake. Events E3 and E4 have not been reported in previous studies and we did not observe event E1 (300-430 cal yr B.P.) at the site. Based on displacement and fault segmentation from the geologic features, we argue that the two new events may have occurred along the northern part of the Chelungpu fault. The vertical slip rate is estimated to be at least 3.9 ‫ע‬ 0.2 mm/yr over the past 2 ka, which is similar to the long-term estimation through a calculation of late Pleistocene-Holocene terrace elevations on the hanging wall.

Biological systems, ranging from bacteria and fungi to humans, can methylate arsenic. Recent studies have suggested that the AsIII S-adenosylmethionine methyltransferase (arsM) gene in bacteria was responsible for the removal of arsenic as the volatile arsines from the bacteria. However, there has been no direct measure of the arsines released from bacteria cultures. We describe here an integrated system incorporating the bacterial incubation and volatile arsenic species analysis, and we demonstrate its application to the identification of the volatile arsines produced in bacterial cultures. The headspace of the bacterial cultures was purged with helium, and the volatile arsenic species were trapped in a chromatographic column immersed in liquid nitrogen. The cryogenically trapped arsines [AsH3, (CH3)AsH2, (CH3)2AsH, and (CH3)3As] were separated by gas chromatography and were detected by inductively coupled plasma mass spectrometry. A hydride generation system was coupled to the bacterial culture system, allowing for spiking standards and for generating calibration arsines necessary for quantitative analysis. Both bacteria containing the arsM gene or its variant arsMC2 gene were able to produce 400-500 ng of trimethylarsine. No trimethylarsine was detectable in bacteria lacking the arsM gene (containing the vector plasmid as negative control). These results confirm that arsM is responsible for releasing arsenic as volatile species from the arsenic-resistant bacteria. Our results also show traces of AsH3, CH3AsH2, and (CH3)2AsH in cultures of bacteria expressing arsM. The method detection limits for AsH3, CH3AsH2, (CH3)2AsH, and (CH3)3As were 0.5, 0.5, 0.7, and 0.6 pg, respectively. The ability to quantify trace levels of these volatile arsenic species makes it possible to study the biotransformation and biochemical roles of the evolution of these volatile arsenic species by biological systems.

[1] Taiwan's 1999 M w 7.6 earthquake generated over 85 km surface rupture along the Chelungpu thrust fault. Paleoseismic studies at the Shi-Jia site near Nantou city, reveal folding as the predominant form of deformation. Stratigraphic relations across the 1999 fold scarp show the style and degree of deformation caused by the penultimate event is similar to observed 1999 deformation. A boring transect across the fold scarp provides additional evidence of an earlier earthquake. Investigations at the Shi-Jia site revealed three prehistoric events; accelerator mass spectrometry (AMS) radiocarbon ages indicate that the penultimate earthquake occurred between 1160 and 1440 A.D. Paleoseismic studies north of the Shi-Jia site reveal much younger penultimate earthquakes, suggesting a 1999-type event may not be characteristic along the Tsaotun segment of the Chelungpu fault.

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