Holocarboxylase Synthetase-dependent Biotinylation of Histone H4

Holocarboxylase synthetase (HCS) catalyzes the binding of biotin to lysine (K) residues in histones H3 and H4. Histone biotinylation marks play important roles in the repression of genes and retrotransposons. Preliminary studies suggested that K16 in histone H4 is a target for biotinylation by HCS. Here we demonstrated that H4K16bio is overrepresented in repeat regions {pericentromeric alpha satellite repeats; long terminal repeats (LTR)} compared with euchromatin promoters. H4K16bio was also enriched in the repressed interleukin-2 gene promoter. The enrichment at LTR22 and promoter 1 of the sodium-dependent multivitamin transporter (SMVT) depended on biotin supply; and was significantly lower in fibroblasts from an HCS-deficient patient compared with an HCS wild-type control. We conclude that H4K16bio is a real phenomenon and plays a role in the transcriptional repression of repeats and genes. HCS catalyzes the covalent binding of biotin to carboxylases, in addition to its role as a histone biotinyl ligase. HCS null individuals are not viable whereas HCS deficiency is linked to developmental delays and phenotypes such as short life span and low stress resistance. Here, we developed a 96-well plate assay for high-throughput analysis of HCS based on the detection of biotinylated p67 using IRDye-streptavidin and infrared spectroscopy. We demonstrated that the catalytic activity of rHCS depends on temperature and time, and proposed optimal substrate and enzyme concentrations to ensure ideal measurement of rHCS activity and its kinetics. Additionally, we demonstrated that this assay is sensitive enough to detect biotinylation of p67 by endogenous HCS from Jurkat lymphoid cells.

High Resolution Paleoceanography of the Guaymas Basin, Gulf of California, During the Past 15 000 Years

Deep Sea Drilling Project Site 480 (27°54.10’N, 111°39.34’W; 655 m water depth) contains a high resolution record of paleoceanographic change of the past 15 000 years for the Guaymas Basin, a region of very high diatom productivity within the central Gulf of California. Analyses of diatoms and silicoflagellates were completed on samples spaced every 40-50 yr, whereas ICP-AES geochemical analyses were completed on alternate samples (sample spacing 80-100 yr). The Bolling-Allerod interval (14.6-12.9 ka) (note, ka refers to 1000 calendar years BP throughout this report) is characterized by an increase in biogenic silica and a decline in calcium carbonate relative to surrounding intervals, suggesting conditions somewhat similar to those of today. The Younger Dryas event (12.9-11.6 ka) is marked by a major drop in biogenic silica and an increase in calcium carbonate. Increasing relative percentage contributions of Azpeitia nodulifera and Dictyocha perlaevis (a tropical diatom and silicoflagellate, respectively) and reduced numbers of the silicoflagellate Octactis pulchra are supportive of reduced upwelling of nutrient-rich waters. Between 10.6 and 10.0 ka, calcium carbonate and A. nodulifera abruptly decline at DSDP 480, while Roperia tesselata, a diatom indicative of winter upwelling in the modern-day Gulf, increases sharply in numbers. A nearly coincident increase in the silicoflagellate Dictyocha stapedia suggests that waters above DSDP 480 were more similar to the cooler and slightly more saline waters of the northern Gulf during much of the early and middle parts of the Holocene (~10 to 3.2 ka). At about 6.2 ka a stepwise increase in biogenic silica and the reappearance of the tropical diatom A. nodulifera marks a major change in oceanographic conditions in the Gulf. A winter shift to more northwesterly winds may have occurred at this time along with the onset of periodic northward excursions (El Nino-driven?) of the North Equatorial Countercurrent during the summer. Beginning between 2.8 and 2.4 ka, the amplitude of biogenic silica and wt% Fe, Al, and Ti (proxies of terrigenous input) increase, possibly reflecting intensification of ENSO cycles and the establishment of modern oceanographic conditions in the Gulf. Increased numbers of O. pulchra after 2.8 ka suggest enhanced spring upwelling.