Systematics and Biogeography of the Red Algal Genus Yonagunia (Halymeniaceae, Rhodophyta) from the Indo-Pacific Including the Description of Two New Species from Taiwan.

Carpopeltis maillardii has been regarded as a widely distributed species in the Indo-Pacific region. In this study, we analyzed the genetic diversity of C. maillardii and related species collected from Taiwan and the Indian Ocean based on rbcL sequences, in order to elucidate species boundaries, diversity, and biogeographic patterns. Our analyses show that C. maillardii specimens are only distantly related to the genus Carpopeltis (type: C. phyllophora) but instead form a clade together with species of Yonagunia. We therefore propose the new combination Yonagunia maillardii comb. nov. In addition, two new species (Yonagunia palmata sp. nov. and Yonagunia taiwani-borealis sp. nov.) are described from Taiwan. The close relationship of Yonagunia to Grateloupia is corroborated by detailed observations of the female reproductive structures, which demonstrate that the development of auxiliary cell ampullae before and after diploidization is similar to that of Grateloupia sensu stricto. Namely, the ampullae are composed of only two orders of unbranched filaments in which only a few ampullar cells are incorporated into a basal fusion cell after diploidization of the auxiliary cell and the pericarp consists almost entirely of secondary medullary filaments. Of all Yonagunia species, Y. maillardii has the widest distribution in the Indo-Pacific, and can be identified in the field by its relatively thin, feathery, and highly branched morphology. Most other species, including those that occur in Taiwan, are seemingly more range-restricted. Our phylogenetic analyses resulted in a well-resolved phylogeny of Yonagunia, with an origin estimated in the Eocene-Oligocene, and diversification of species mainly in the Miocene.

Design, synthesis, and antimicrobial activities of novel functional peptides against Gram-positive and Gram-negative bacteria.

The extensive use of antibiotics in medicine results in the multidrug resistance of bacteria, making the development of new antimicrobial agents an urgent need. Antimicrobial peptides (AMPs) are considered as a new class of antibiotic with characteristics including an ability to kill target cells rapidly and a broad spectrum of activity. We have developed a potent antimicrobial peptide MAP-0403 (MIC = 5 µM), but it exhibited a high hemolytic side-effect (70.7%). To reduce its hemolytic effect and increase antimicrobial activity, three peptides derivatives of MAP-0403 (J-1, J-2, and J-3) were designed, synthesized by solid phase synthesis, purified by RP-HPLC, and characterized by MALDI-TOF MS. Structure-activity relationships of these peptides were studied by using circular dichroism and antimicrobial assays. The percentage of helical structure in J-1, J-2, and J-3 was lower than that of MAP-0403. The antimicrobial activity of J-1 was the same as that of MAP-0403 (MIC = 5 µM), J-2 exhibited the highest activity (MIC = 2.5 µM), while J-3 showed decreased activity (MIC = 10 µM). Compared to MAP-0403, J-2 showed significantly reduced hemolytic effect (3.4%), while J-1 and J-3 showed slightly decreased hemolytic effect (46.2%, 55.6%, respectively). Peptide J-2 was discovered as a novel and potent antimicrobial agents.

Untying a Protein Knot by Circular Permutation.

Topologically knotted proteins are tantalizing examples of how polypeptide chains can explore complex free energy landscapes to efficiently attain defined knotted conformations. The evolution trails of protein knots, however, remain elusive. We used circular permutation to change an evolutionally conserved topologically knotted SPOUT RNA methyltransferase into an unknotted form. The unknotted variant adopted the same three-dimensional structure and oligomeric state as its knotted parent, but its folding stability was markedly reduced with accelerated folding kinetics and its ligand binding was abrogated. Our findings support the hypothesis that the universally conserved knotted topology of the SPOUT superfamily evolved from unknotted forms through circular permutation under selection pressure for folding robustness and, more importantly, for functional requirements associated with the knotted structural element.

WSSV-induced crayfish Dscam shows durable immune behavior.

One of the major gaps in our understanding of arthropod specific immune priming concerns the mechanism[s] by which the observed long-term (>2 weeks) protective effects might be mediated. Hypervariable Dscam (Down syndrome cell adhesion molecule) might support arthropod innate immunity with specificity for more extended periods. We show here that, in the relatively long-lived arthropod Cherax quadricarinatus, CqDscam does not behave like a typical, immediately-acting, short-lived innate immune factor: CqDscam was not induced within hours after challenge with a lethal virus, but instead was only up-regulated after 2-5 days. This initial response faded within ∼ 2 weeks, but another maximum was reached ∼ 1 month later. At around 2 months after the initial challenge, the virus-induced CqDscam bound to the virus virion and acted to neutralize the virus However, although CqDscam helped crayfish to survive during persistent infection, it nevertheless failed to provide any enhanced protection against a subsequent WSSV challenge. Thus, CqDscam is capable of supporting extended anti-virus immune memory in arthropods. Also, during a persistent virus infection, the balance of "immune firepower" in crayfish appears to be altered such that the general immune factors become depleted while CqDscam becomes relatively predominant.

Properties of Litopenaeus vannamei Dscam (LvDscam) isoforms related to specific pathogen recognition.

Arthropod Down syndrome cell adhesion molecules (Dscam) may sometimes function as hypervariable pathogen recognition receptors. They consist of an extracellular region and a cytoplasmic tail, both of which are highly variable. In shrimp, tail-less Dscam proteins (Dscams) have recently been identified, and these appear to be unlike other arthropod extracellular Dscams that are released from the cell membrane by proteolytic cleavage. Here we investigate the properties of these unique shrimp proteins and show that they can be directly secreted from transfected cells. We also investigate the diverse cytoplasmic tail variants of membrane-bound shrimp Dscams, and show that elements E1A and E3 seem to be related to Dscam immune function. Challenge with Vibrio harveyi not only enhanced total Dscam and the immune-related cytoplasmic tail variants, but also induced expression of certain Ig2 + Ig3 combinations. A pathogen binding assay with these Ig2 + Ig3 extracellular variants showed that both the V. harveyi-induced Dscams and Dscams induced by buffer injection could be either pathogen-specific or specific only for Gram-negative pathogens, while other "general" Dscam variants were sensitive to a wide range of pathogens. The same assay also suggested that shrimp Dscam isoforms show a stronger response to the host's natural pathogens.