Holliday junction resolvase in Schizosaccharomyces pombe has identical endonuclease activity to the CCE1 homologue YDC2

A novel Holliday junction resolving activity has been identified in fractionated cell extracts of the fission yeast Schizosaccharomyces pombe . The enzyme catalyses endonucleolytic cleavage of Holliday junction-containing chi DNA and synthetic four-way DNA junctions. The activity cuts with high specificity a synthetic four-way junction containing a 12 bp core of homologous sequences but has no activity on another four-way junction (with a fixed crossover point), a three-way junction, linear duplex DNA or duplex DNA containing six mismatched nucleotides in the centre. The major cleavage sites map as single nicks in the vicinity of the crossover point, 3' of a thymidine residue. These data indicate that the activity has a strong DNA structure selectivity as well as a limited sequence preference; features similar to the Holliday junction resolving enzymes RuvC of Escherichia coli and the mitochondrial CCE1 (cruciform-cuttingenzyme 1) of Saccharomyces cerevisiae. A putative homologue of CCE1 in S.pombe (YDC2_SCHPO) has been identified through a search of the sequence database. The open reading frame of this gene has been cloned and the encoded protein, YDC2, expressed in E.coli . The purified recombinant YDC2 exhibits Holliday junction resolvase activity and is, therefore, a functional S.pombe homologue of CCE1. The resolvase YDC2 shows the same substrate specificity and produces identical cleavage sites as the activity obtained from S. pombe cells. Both YDC2 and the cellular activity cleave Holliday junctions in both orientations to give nicks that can be ligated in vitro. The partially purified Holliday junction resolving enzyme in fission yeast is biochemically indistinguishable from recombinant YDC2 and appears to be the same protein

Improved recovery from prolonged exercise following the consumption of low glycemic index carbohydrate meals

The present study examined the effects of the glycemic index (GI) of post-exercise carbohydrate (CHO) intake on endurance capacity and the metabolic responses during exercise the following day. Nine active males participated in two trials in a randomised crossover design. The experimental protocol was completed over two days. On day 1, subjects completed a 90 min treadmill run at 70% VO2 max (R1). Thereafter, they were supplied with a diet consisting of either high glycemic index (HGI) or low glycemic index (LGI) CHO and provided 8g CHOkg body mass (BM)-1.On day 2, after an overnight fast, subjects ran to exhaustion at 70% VO2 max (R2). Eight subjects completed both performance runs (R2). Run time to exhaustion during R2 was longer in the LGI trial (108.9 7.4 min) than in the HGI trial (96.9 4.8 min) (p<0.05). Average RER values were higher in the HGI trial compared to the LGI trial (p<0.05). Fat oxidation rates and free fatty acid concentrations were higher in the LGI trial than the HGI trial (p<0.05). The results of the study suggest that the increased endurance capacity during R2 was largely a consequence of the greater fat oxidation following the consumption of the LGI meals

The spatial variations of lightning during small Florida thunderstorms

Networks of field mills (FM's) and lightning direction finders (LDF's) were used to locate lightning over the NASA KSC on three storm days. Over 90 percent of all cloud-to-ground (CG) flashes that were detected by the LDF's in the study area were also detected by the LDF's. About 17 percent of the FM CG events could be fitted to either a monopole or a dipole charge model. These projected FM charge locations are compared to LDF locations, i.e., the ground strike points. It was found that 95 percent of the LDF points are within 12 km of the FM charge, 75 percent are within 8 km, and 50 percent are within 4 km. For a storm on 22 Jul. 1988, there was a systematic 5.6 km shift between the FM charge centers and the LDF strike points that might have been caused by the meteorological structure of the storm

Type 1 diabetes

Type 1 diabetes is a chronic autoimmune disease characterised by insulin deficiency and resultant hyperglycaemia. Knowledge of type 1 diabetes has rapidly increased over the past 25 years, resulting in a broad understanding about many aspects of the disease, including its genetics, epidemiology, immune and β-cell phenotypes, and disease burden. Interventions to preserve β cells have been tested, and several methods to improve clinical disease management have been assessed. However, wide gaps still exist in our understanding of type 1 diabetes and our ability to standardise clinical care and decrease disease-associated complications and burden. This Seminar gives an overview of the current understanding of the disease and potential future directions for research and care