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.

FOREST ANIMAL DAMAGE CONTROL

As a nation we have gained world recognition for our ability to utilize our resources. In forestry our greatest accomplishments have been in the mechanization of harvest methods and in improvements in forest products. The renewal of this resource has been our greatest neglect. Though the end of the 19th Century marked the beginning of the conservation movement, it was not until a half century later that the force of economics through the demands of a growing population made forest re-establishment more than just a desire. Conservation in itself is a Utopian concept which requires other motivating forces to make it a reality. In the post-war years, and as late as the early 195O's, stocked land in the Pacific Northwest could be purchased for less than the cost of planting; the economic incentive was lacking. Only with sustained yield management and increased land values was there a balance in favor of true values. With greater effort placed on forest regeneration there was an increased need for methods of reducing losses to wildlife. The history of forest wildlife damage research, therefore, parallels that of forest land management; after rather austere beginnings, development became predominantly a response to economics. It was not until 1950 that the full time of one scientist was assigned to this important activity. The development of control methods for forest animal damage is a relatively new area of research. All animal life is dependent upon plants for its existence; forest wildlife is no exception. The removal of seed and foliage of undesirable plants often benefits the land managers; only when the losses or injuries are in conflict with man's interest is there damage involved. Unfortunately, the feeding activities of wildlife and the interests of the land managers are often in conflict. Few realize the breadth, scope, and subtilities associated with forest wildlife damage problems. There are not only numerous species of animals involved, but also a myriad of conditions, each combination possessing unique facets. It is a foregone conclusion that an understanding of the conditions is essential to facilitate a solution to any given problem. Though there are numerous methods of reducing animal damage, all of which have application under some situations, in this discussion emphasis will be placed on the role of chemicals and on western problems. Because of the broadness and complexity of the problem, generalizing is necessary and only brief coverage will be possible. However, an attempt will be made to discuss the use and limitations of various control methods.