Mechanism of RAD51-dependent DNA interstrand cross-link repair

DNA interstrand cross-links (ICLs) are toxic DNA lesions whose repair in S phase of eukaryotic cells is incompletely understood. In Xenopus egg extracts, ICL repair is initiated when two replication forks converge on the lesion. Dual incisions then create a DNA double-strand break (DSB) in one sister chromatid, whereas lesion bypass restores the other sister. We report that the broken sister chromatid is repaired via RAD51-dependent strand invasion into the regenerated sister. Recombination acts downstream of FANCI-FANCD2, yet RAD51 binds ICL-stalled replication forks independently of FANCI-FANCD2 and before DSB formation. Our results elucidate the functional link between the Fanconi anemia pathway and the recombination machinery during ICL repair. In addition, they demonstrate the complete repair of a DSB via homologous recombination in vitro.     

Structure and mechanism of the lactose permease of Escherichia coli

Membrane transport proteins that transduce free energy stored in electrochemical ion gradients into a concentration gradient are a major class of membrane proteins. We report the crystal structure at 3.5 angstroms of the Escherichia coli lactose permease, an intensively studied member of the major facilitator superfamily of transporters. The molecule is composed of N- and C-terminal domains, each with six transmembrane helices, symmetrically positioned within the permease. A large internal hydrophilic cavity open to the cytoplasmic side represents the inward-facing conformation of the transporter. The structure with a bound lactose homolog, beta-D-galactopyranosyl-1-thio-beta-D-galactopyranoside, reveals the sugar-binding site in the cavity, and residues that play major roles in substrate recognition and proton translocation are identified. We propose a possible mechanism for lactose/proton symport (co-transport) consistent with both the structure and a large body of experimental data.     

A distinct small RNA pathway silences selfish genetic elements in the germline

In the Drosophila germline, repeat-associated small interfering RNAs (rasiRNAs) ensure genomic stability by silencing endogenous selfish genetic elements such as retrotransposons and repetitive sequences. Whereas small interfering RNAs (siRNAs) derive from both the sense and antisense strands of their double-stranded RNA precursors, rasiRNAs arise mainly from the antisense strand. rasiRNA production appears not to require Dicer-1, which makes microRNAs (miRNAs), or Dicer-2, which makes siRNAs, and rasiRNAs lack the 2',3' hydroxy termini characteristic of animal siRNA and miRNA. Unlike siRNAs and miRNAs, rasiRNAs function through the Piwi, rather than the Ago, Argonaute protein subfamily. Our data suggest that rasiRNAs protect the fly germline through a silencing mechanism distinct from both the miRNA and RNA interference pathways.     

Evaluation of BAM (butorphanol–azaperone–medetomidine) in captive African lion (Panthera leo) immobilization

Objective

The combination of butorphanol, azaperone and medetomidine (BAM) with subsequent antagonism by naltrexone–yohimbine or naltrexone–atipamezole was evaluated for reversible immobilization of captive African lions (Panthea leo).

‘Relationships between relationships’ in forest stands: intercepts and exponents analyses

Relationships between diameter at breast height(dbh) versus stand density, and tree height versus dbh(height curve) were explored with the aim to find if there were functional links between correspondent parameters of the relationships, exponents and intercepts of their power functions. A geometric model of a forest stand using a conic approximation suggested that there should be interrelations between correspondent exponents and intercepts of the relationships. It is equivalent to a type of ‘relationship between relationships’ that might exist in a forest stand undergoing self-thinning, and means that parameters of one relationship may be predicted from parameters of another. The predictions of the model were tested with data on forest stand structure from published databases that involved a number of trees species and site quality levels. It was found that the correspondent exponents and intercepts may be directly recalculated from one another for the simplest case when the total stem surface area was independent of stand density. For cases where total stem surface area changes with the drop of density, it is possible to develop a generalization of the model in which the interrelationships between correspondent parameters(exponents and intercepts) may be still established.     

Effects of early leaf removal on grape yield,chemical characteristics,and antioxidant activity of grape variety Cabernet Sauvignon and wine from eastern Croatia

The aim of this study was to determine the impact of two different treatments of early defoliation performed before blooming on: grape yield, chemical parameters, polyphenols content, and antioxidant activity of grape and red wine cv. Cabernet Sauvignon from the vineyard located in Ilok, the eastern continental region of Croatia. Two different treatments of leaf removal (LR) were performed: removal of 3 leafs (T1) and 6 leafs (T2) before blooming, together with control (no leaf removal) (T3) during two years (2013 and 2014). Crop yield and average cluster weights per vine were determined. Density, pH and titratable acidity were measured in must, while the total phenols, total anthocyanins and antioxidant activity were measured in the extract of grape skin and produced wine. The analysis of individual anthocyanins in wine was performed by HPLC method. T2 treatment significantly lowered the crop yield and the average cluster weights, and increased total phenols, total anthocyanins, antioxidant activity and most abundant individual anthocyanins in wine. Defoliation did not affect the other chemical parameters in must, grape skin extract and wine. Vintage year is statistically the most significant source of variability for density of must, antioxidant activity in grape skin extract, as well as pH and titratable acidity in wine. This study has showed that the early leaf removal treatment in eastern continental part of Croatia could be used for the production of smaller quantity of high quality Cabernet Sauvignon red wine abundant with anthocyanins.