Characterisation and engineering of a thermophilic RNA ligase from Palaeococcus pacificus.

RNA ligases are important enzymes in molecular biology and are highly useful for the manipulation and analysis of nucleic acids, including adapter ligation in next-generation sequencing of microRNAs. Thermophilic RNA ligases belonging to the RNA ligase 3 family are gaining attention for their use in molecular biology, for example a thermophilic RNA ligase from Methanobacterium thermoautotrophicum is commercially available for the adenylation of nucleic acids. Here we extensively characterise a newly identified RNA ligase from the thermophilic archaeon Palaeococcus pacificus (PpaRnl). PpaRnl exhibited significant substrate adenylation activity but low ligation activity across a range of oligonucleotide substrates. Mutation of Lys92 in motif I to alanine, resulted in an enzyme that lacked adenylation activity, but demonstrated improved ligation activity with pre-adenylated substrates (ATP-independent ligation). Subsequent structural characterisation revealed that in this mutant enzyme Lys238 was found in two alternate positions for coordination of the phosphate tail of ATP. In contrast mutation of Lys238 in motif V to glycine via structure-guided engineering enhanced ATP-dependent ligation activity via an arginine residue compensating for the absence of Lys238. Ligation activity for both mutations was higher than the wild-type, with activity observed across a range of oligonucleotide substrates with varying sequence and secondary structure.

Evaluating the attractiveness and effectiveness of artificial coral reefs as a recreational ecosystem service.

Artificial reefs are increasingly being used around the globe to attract recreational divers, for both environmental and commercial reasons. This paper examines artificial coral reefs as recreational ecosystem services (RES) by evaluating their attractiveness and effectiveness and by examining divers' attitudes toward them. An online survey targeted at divers in Israel (n = 263) indicated that 35% of the dives in Eilat (a resort city on the shore of the Red Sea) take place at artificial reefs. A second study monitored divers' behavior around the Tamar artificial reef, one of the most popular submerged artificial reefs in Eilat, and juxtaposed it with divers' activities around two adjacent natural reefs. Findings show that the average diver density at the artificial reef was higher than at the two nearby natural knolls and that the artificial reef effectively diverts divers from natural knolls. A third study that examined the attitudes towards natural vs. artificial reefs found that the artificial reefs are considered more appropriate for training, but that divers feel less relaxed around them. By utilizing the RES approach as a framework, the study offers a comprehensive methodology that brings together the aesthetic, behavioral, and attitudinal aspects in terms of which artificial reefs can be evaluated.

Size Matters: Observed and Modeled Camouflage Response of European Cuttlefish (Sepia officinalis) to Different Substrate Patch Sizes during Movement.

Camouflage is common throughout the phylogenetic tree and is largely used to minimize detection by predator or prey. Cephalopods, and in particular Sepia officinalis cuttlefish, are common models for camouflage studies. Predator avoidance behavior is particularly important in this group of soft-bodied animals that lack significant physical defenses. While previous studies have suggested that immobile cephalopods selectively camouflage to objects in their immediate surroundings, the camouflage characteristics of cuttlefish during movement are largely unknown. In a heterogenic environment, the visual background and substrate feature changes quickly as the animal swim across it, wherein substrate patch is a distinctive and high contrast patch of substrate in the animal's trajectory. In the current study, we examine the effect of substrate patch size on cuttlefish camouflage, and specifically the minimal size of an object for eliciting intensity matching response while moving. Our results indicated that substrate patch size has a positive effect on animal's reflectance change, and that the threshold patch size resulting in camouflage response falls between 10 and 19 cm (width). These observations suggest that the animal's length (7.2-12.3 cm mantle length in our case) serves as a possible threshold filter below which objects are considered irrelevant for camouflage, reducing the frequency of reflectance changes-which may lead to detection. Accordingly, we have constructed a computational model capturing the main features of the observed camouflaging behavior, provided for cephalopod camouflage during movement.