Ages and tephra layers in Chatham Rise sediment cores

We present sea surface temperature (SST) estimates based on the relative abundances of long-chain C37 alkenones (UK37') in four sediment cores from a transect spanning the subtropical to subantarctic waters across the subtropical front east of New Zealand. SST estimates from UK37' are compared to those derived from foraminiferal assemblages (using the modern analog technique) in two of these cores. Reconstructions of SST in core tops and Holocene sediments agree well with modern average summer temperatures of ~18°C in subtropical waters and ~14°C in subpolar waters, with a 4°-5°C gradient across the front. Down core UK37' SST estimates indicate that the regional summer SST was 4°-5°C cooler during the last glaciation with an SST of ~10°C in subpolar waters and an SST of ~14°C in subtropical waters. Temperature reconstructions from foraminiferal assemblages agree with those derived from alkenones for the Holocene. In subtropical waters, reconstructions also agree with a glacial cooling of 4° to ~14°C. In contrast, reconstructions for subantarctic pre-Holocene waters indicate a cooling of 8°C with glacial age warm season water temperatures of ~6°C. Thus the alkenones suggest the glacial temperature gradient across the front was the same or reduced slightly to 3.5°-4°C, whereas foraminiferal reconstructions suggest it doubled to 8°C. Our results support previous work indicating that the STF remained fixed over the Chatham Rise during the Last Glacial Maximum. However, the differing results from the two techniques require additional explanation. A change in euphotic zone temperature profiles, seasonality of growth, or preferred growth depth must have affected the temperatures recorded by these biologically based proxies. Regardless of the specific reason, a differential response to the environmental changes between the two climate regimes by the organisms on which the estimates are based suggests increased upwelling associated with increased winds and/or a shallowing of the thermocline associated with increased stratification of the surface layer in the last glaciation.

Can Jupiters be found by monitoring Galactic bulge microlensing events from northern sites?

In 1998 the EXPORT team monitored microlensing event light curves using a charge-coupled device (CCD) camera on the IAC 0.8-m telescope on Tenerife to evaluate the prospect of using northern telescopes to find microlens anomalies that reveal planets orbiting the lens stars. The high airmass and more limited time available for observations of Galactic bulge sources make a northern site less favourable for microlensing planet searches. However, there are potentially a large number of northern 1-m class telescopes that could devote a few hours per night to monitor ongoing microlensing events. Our IAC observations indicate that accuracies sufficient to detect planets can be achieved despite the higher airmass.

Direct isolation of human transcribed sequences from yeast artificial chromosomes through the application of RNA fingerprinting

The identification of cDNA clones from genomic regions known to contain human genes is usually the rate-limiting factor in positional cloning strategies. We demonstrate here that human genes present on yeast artificial chromosomes (YACs) are transcribed in yeast host cells. We have used the arbitrarily primed RNA (RAP) fingerprinting method to identify human-specific, transcribed sequences from YACs located in the 13q12 chromosome region. By comparing the RAP fingerprints generated using defined, arbitrary primers from various fragmented YACs, megaYACs, and host yeast, we were able to identify and map 20 products transcribed from the human YAC inserts. This method, therefore, permits the simultaneous isolation and mapping of novel expressed sequences directly from whole YACs.

The helical domain of a G protein α subunit is a regulator of its effector

The α subunit (Gα) of heterotrimeric G proteins is a major determinant of signaling selectivity. The Gα structure essentially comprises a GTPase “Ras-like” domain (RasD) and a unique α-helical domain (HD). We used the vertebrate phototransduction model to test for potential functions of HD and found that the HD of the retinal transducin Gα (Gαt) and the closely related gustducin (Gαg), but not Gαi1, Gαs, or Gαq synergistically enhance guanosine 5′-γ[-thio]triphosphate bound Gαt (GαtGTPγS) activation of bovine rod cGMP phosphodiesterase (PDE). In addition, both HDt and HDg, but not HDi1, HDs, or HDq attenuate the trypsin-activated PDE. GαtGDP and HDt attenuation of trypsin-activated PDE saturate with similar affinities and to an identical 38% of initial activity. These data suggest that interaction of intact Gαt with the PDE catalytic core may be caused by the HD moiety, and they indicate an independent site(s) for the HD moiety of Gαt within the PDE catalytic core in addition to the sites for the inhibitory Pγ subunits. The HD moiety of GαtGDP is an attenuator of the activated catalytic core, whereas in the presence of activated GαtGTPγS the independently expressed HDt is a potent synergist. Rhodopsin catalysis of Gαt activation enhances the PDE activation produced by subsaturating levels of Gαt, suggesting a HD-moiety synergism from a transient conformation of Gαt. These results establish HD-selective regulations of vertebrate retinal PDE, and they provide evidence demonstrating that the HD is a modulatory domain. We suggest that the HD works in concert with the RasD, enhancing the efficiency of G protein signaling.

Reducing the flexibility of retinal restores a wild-type-like photocycle in bacteriorhodopsin mutants defective in protein–retinal coupling

The thermal re-isomerization of retinal from the 13-cis to the all-trans state is a key step in the final stages of the photocycle of the light-driven proton pump, bacteriorhodopsin. This step is greatly slowed upon replacement of Leu-93, a residue in van der Waals contact with retinal. The most likely role of this key interaction is that it restricts the flexibility of retinal. To test this hypothesis, we have exchanged native retinal in Leu-93 mutants with bridged retinal analogs that render retinal less flexible by restricting free rotation around either the C10—C11 (9,11-bridged retinal) or C12—C13 (11,13-bridged retinal) single bonds. The effect of the analogs on the photocycle was then determined spectroscopically by taking advantage of the previous finding that the decay of the O intermediate in the Leu-93 mutants provides a convenient marker for retinal re-isomerization. Time-resolved spectroscopic studies showed that both retinal analogs resulted in a dramatic acceleration of the photocycling time by increasing the rate of decay of the O intermediate. In particular, exchange of native retinal in the Leu-93 → Ala mutant with the 9,11-bridged retinal resulted in an acceleration of the decay of the O intermediate to a rate similar to that seen in wild-type bacteriorhodopsin. We conclude that the protein-induced restriction of conformational flexibility in retinal is a key structural requirement for efficient protein–retinal coupling in the bacteriorhodopsin photocycle.