DNA alignment and Bayesian trees of partial sequences of COI, COIII and the haemocyanin functional unit f-g of 28 octopod species

Background: Octopods have successfully colonised the world's oceans from the tropics to the poles. Yet, successful persistence in these habitats has required adaptations of their advanced physiological apparatus to compensate impaired oxygen supply. Their oxygen transporter haemocyanin plays a major role in cold tolerance and accordingly has undergone functional modifications to sustain oxygen release at sub-zero temperatures. However, it remains unknown how molecular properties evolved to explain the observed functional adaptations. We thus aimed to assess whether natural selection affected molecular and structural properties of haemocyanin that explains temperature adaptation in octopods. Results: Analysis of 239 partial sequences of the haemocyanin functional units (FU) f and g of 28 octopod species of polar, temperate, subtropical and tropical origin revealed natural selection was acting primarily on charge properties of surface residues. Polar octopods contained haemocyanins with higher net surface charge due to decreased glutamic acid content and higher numbers of basic amino acids. Within the analysed partial sequences, positive selection was present at site 2545, positioned between the active copper binding centre and the FU g surface. At this site, methionine was the dominant amino acid in polar octopods and leucine was dominant in tropical octopods. Sites directly involved in oxygen binding or quaternary interactions were highly conserved within the analysed sequence. Conclusions: This study has provided the first insight into molecular and structural mechanisms that have enabled octopods to sustain oxygen supply from polar to tropical conditions. Our findings imply modulation of oxygen binding via charge-charge interaction at the protein surface, which stabilize quaternary interactions among functional units to reduce detrimental effects of high pH on venous oxygen release. Of the observed partial haemocyanin sequence, residue 2545 formed a close link between the FU g surface and the active centre, suggesting a role as allosteric binding site. The prevalence of methionine at this site in polar octopods, implies regulation of oxygen affinity via increased sensitivity to allosteric metal binding. High sequence conservation of sites directly involved in oxygen binding indicates that functional modifications of octopod haemocyanin rather occur via more subtle mechanisms, as observed in this study.

Annotated record of the detailed examination of Mn deposits from DSDP Hole 2-8

The Atlantic Advisory Panel proposed that Site 8 should be drilled on the rise between the Hatteras and Sohm Abyssal Plains (lat 35° 2l'N., long 67° 3l'W.) This location was considered to offer the best opportunity for realizing two primary objectives. The first of these objectives was to sample and date the oldest available rock in a region adjacent to the North American continent and as far as possible from the Mid-Atlantic Ridge. The second objective relates to the potential paleobiological and paleoecological information that could be derived from the sedimentary column in this general area.

Annotated record of the detailed examination of Mn deposits from DSDP Hole 2-12C

Site 12 of this report formerly was designated 14 by the Atlantic Advisory Panel. It was drilled in the Cape Verde Basin (latitude 19° 42'N, longitude 26° 01'W), which is underlain by 2000 to 2100 feet (0.9 to 1.1 seconds reflection time) of sediment. The bottom topography of the basin is fairly smooth but not to the degree of an abyssal plain. Site 12 was selected to provide material for paleontological investigations.

Annotated record of the detailed examination of Mn deposits from DSDP Hole 2-9A

The Atlantic Advisory Panel proposed that Site 9 should be drilled on the northeastern flank of the Bermuda Rise (lat. 32° 37' N., long. 59° 10' W.), which is about 100 miles west of the Sohm Abyssal Plain. The bottom of this region consists of low linear ridges that are roughly parallel and oriented in a northwest-southeasterly direction. Scattered seamounts, some of which have peaks 2000 fathoms (3660 meters) below sea level, arise from the otherwise featureless sea floor between the ridges. The primary purpose in drilling Site 9 was to examine a sedimentary column where seismic reflectors were largely absent and to determine the age of sediments overlying acoustical basement in the examination of sea floor spreading.