Review and phylogeny of the New Zealand hagfishes (Myxiniformes: Myxinidae), with a description of three new species

Hagfishes from New Zealand are reviewed and a phylogeny proposed using morphological and genetic data (DNA sequences of cytochrome c oxidase subunit I gene, COI, and the small subunit RNA, 16S). E ptatretus cryptus sp. nov. was previously confused with Eptatretus cirrhatus (Forster in Bloch & Schneider, 1801) because of their similar morphology, and is found from the Three Kings Islands to Stewart Island and in the eastern part of the Chatham Rise (at depths of 96–922 m). E ptatretus poicilus sp. nov. is endemic to the Three Kings Islands, where it is common and associated with soft sediment and deep‐sea coral‐sponge habitats (114–842 m). N eomyxine caesiovitta sp. nov. is a slender hagfish found along the east coast of the North Island south to the Chatham Rise (430–1083 m). A neotype is erected for E. cirrhatus (type locality: Breaksea Sound, Fiordland), occurring widely in New Zealand coastal, shelf, and slope waters (1–922 m), but not at the Three Kings Islands. Eptatetrus goliath Mincarone & Stewart, 2006, Neomyxine biniplicata (Richardson & Jowett, 1951), and Nemamyxine elongata Richardson, 1958 are further described using additional material. Rubicundus eos (Fernholm, 1991) is still only known from the holotype (type locality: Challenger Plateau). Genetic results showed that the New Zealand Eptatretus species form a monophyletic group within the subfamily Eptatretinae, indicating likely speciation from a single common ancestor within the area. E ptatretus poicilus sp. nov. is the sister species of E. cirrhatus, and E . cryptus sp. nov. is closely associated with the clade formed by these two species. Eptatretus goliath is most closely associated with Eptatretus minor Fernholm & Hubbs, 1981 (Gulf of Mexico), these two species basally diverging within New Zealand hagfishes. The endemic genus Neomyxine forms a well‐supported monophyletic group of as yet uncertain position within the phylogenetic tree. A key to the New Zealand hagfishes, fresh colour photographs, distribution maps, and in situ video recordings are presented. © 2015 The Linnean Society of London     

Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail

A phylogenetically conserved RNA structure within the NS5B coding region of hepatitis C virus functions as a cis-replicating element (CRE). Integrity of this CRE, designated SL9266 (alternatively 5BSL3.2), is critical for genome replication. SL9266 forms the core of an extended pseudoknot, designated SL9266/PK, involving long distance RNA–RNA interactions between unpaired loops of SL9266 and distal regions of the genome. Previous studies demonstrated that SL9266/PK is dynamic, with ‘open’ and ‘closed’ conformations predicted to have distinct functions during virus replication. Using a combination of site-directed mutagenesis and locked nucleic acids (LNA) complementary to defined domains of SL9266 and its interacting regions, we have explored the influence of this structure on genome translation and replication. We demonstrate that LNAs which block formation of the closed conformation inhibit genome translation. Inhibition was at least partly independent of the initiation mechanism, whether driven by homologous or heterologous internal ribosome entry sites or from a capped message. Provision of SL9266/PK in trans relieved translational inhibition, and mutational analysis implied a mechanism in which the closed conformation recruits a cellular factor that would otherwise suppresses translation. We propose that SL9266/PK functions as a temporal switch, modulating the mutually incompatible processes of translation and replication.     

Diaryl substituted pyrazoles as potent CCR2 receptor antagonists

We have identified and synthesized a series of diaryl substituted pyrazoles as potent antagonists of the chemokine receptor subtype 2. Structure-activity relationship studies directed toward improving the potency led to the discovery of 23 (IC50 = 6 nM).     

Tertiary interactions between the fifth and sixth transmembrane segments of rhodopsin.

We have used cysteine scanning mutagenesis and disulfide cross-linking in a split rhodopsin construct to investigate the secondary structure and tertiary contacts of the fifth (TM5) and sixth (TM6) transmembrane segments of rhodopsin. Using a simple increase in pH to promote disulfide bond formation, three cross-links between residues on the extracellular side of TM5 (at positions 198, 200, and 204) and TM6 (at position 276) have been identified and characterized. The helical pattern of cross-linking observed indicates that the fifth transmembrane helix extends through residue 200 but does not include residue 198. Rhodopsin mutants containing these disulfides demonstrate nativelike absorption spectra and light-dependent activation of transducin, suggesting that large movements on the extracellular side of TM5 with respect to TM6 are not required for receptor activation.