Role of histone H3 lysine 9 methylation in epigenetic control of heterochromatin assembly

The assembly of higher order chromatin structures has been linked to the covalent modifications of histone tails. We provide in vivo evidence that lysine 9 of histone H3 (H3 Lys9) is preferentially methylated by the Clr4 protein at heterochromatin-associated regions in fission yeast. Both the conserved chromo- and SET domains of Clr4 are required for H3 Lys9 methylation in vivo. Localization of Swi6, a homolog of Drosophila HP1, to heterochomatic regions is dependent on H3 Lys9 methylation. Moreover, an H3-specific deacetylase Clr3 and a beta-propeller domain protein Rik1 are required for H3 Lys9 methylation by Clr4 and Swi6 localization. These data define a conserved pathway wherein sequential histone modifications establish a "histone code" essential for the epigenetic inheritance of heterochromatin assembly.     

Introduction of nucleophiles and spectroscopic probes into antibody combining sites

A general chemical strategy has been developed whereby antibody combining sites can be selectively derivatized with natural or synthetic molecules, such as catalytic groups, drugs, metals, or reporter molecules. Cleavable affinity labels were used to selectively introduce a thiol into the combining site of the immunoglobulin A MOPC 315. This thiol acted both as a nucleophile to accelerate ester thiolysis 60,000-fold and as a handle for selectively derivatizing the antibody with additional functional groups. For example, derivatization of the antibody with a fluorophore made possible a direct spectroscopic assay of antibody-ligand complexation. This chemistry should not only extend our ability to exploit antibody specificity in chemical catalysis, diagnostics, and therapeutics, but may also prove generally applicable to the functional modification of other proteins for which detailed structural information is unavailable.     

Analysis of antibody response by temperature-sensitive measles vaccine strain in the cotton rat model

Measles virus (MeV) vaccine strain, AIK-C, is temperature sensitive (ts), which is thought to be associated with attenuation of virus pathogenicity. In this study, replication and antibody response were examined in cotton rats using viruses carrying different forms of the P gene, which is responsible for the ts phenotype of strain AIK-C and its parental Edmonston strain. When cotton rats were inoculated intranasally, ts viruses neither replicated in lungs, nor reproducibly generated an antibody response. When inoculated intramusculary (i.m.), however, ts strains raised an antibody titer in all animals. This response was not observed when ultraviolet-inactivated virus was used. ts virus, inoculated i.m., was recovered from cotton rat drainage lymph nodes. These results suggest that ts virus, inoculated i.m., could replicate in the cotton rat, presumably at the superficial lymph node, and induce an antibody response. Therefore, cotton rats can serve as a small-animal model for investigating immune responses to safer ts vaccine, as well as recombinant vaccine using AIK-C as a vector for protection against other infectious agents.     

Trickle irrigation: Predominant bacteria in treated Colorado River water and biologically clogged emitters

Summary Bacterial numbers and predominant bacterial types were determined in trickle irrigation systems receiving treated Colorado River water. Fourteen bacterial genera were isolated and identified from 86 water and sediment samples collected from trickle emitter systems receiving six water treatments. The bacteria identified were common aquatic and soil microbes and the genera in order of prevalence were Pseudomonas, Flavobacterium, Vibrio, Brevibacterium, Micrococcus, and Bacillus. A greater diversity of bacterial types was found in water that was sand filtered and received no chemical treatment. Regardless of the water treatment, Pseudomonas stutzeri was the predominant bacterium followed by Flavobacterium lutescens. The data indicated that pigmented bacteria, F. lutescens and ytophaga hutchinsonii caused the yellow color of the slime deposits in biologically clogged emitters; and, their growth was presumably supported by P. stutzeri., a nonpigmented bacterium. The occurrence of Bacillus sp. was enhanced by sand and screen filtration and markedly reduced by chemical conditioning the water with either chlorine and acid or acid alone. No strictly anaerobic bacteria, such as Clostridium sp., were detected in water treated with chlorine and acid. Iron bacteria, Sphaerotilus spp., were not detected in any water or sediment samples from trickle irrigation systems.Contribution from the U.S. Department of Agriculture, Science and Education Administration, Agricultural Research     

Antigenic differences of the scuticociliate Miamiensis avidus from Japan

In Japan and Korea, outbreaks of scuticociliatosis have frequently occurred in Japanese flounder, Paralichthys olivaceus . Morphological observations and small subunit rRNA gene sequences have shown that the causative agent of scuticociliatosis in the flounder is Miamiensis avidus (syn. Philasterides dicentrarchi ). In this study, we elucidated the antigenic differences between six Japanese M. avidus isolates as an initial step toward developing an effective vaccine against the disease. Four Japanese flounder isolates (IyoI, Nakajima, JF05To and Mie0301 isolates), one spotted knifejaw, Oplegnathus punctatus, isolate (SK05Kyo), and one ridged-eye flounder, Pleuronichthys cornutus , isolate (RF05To) were subjected to serological analysis. Antisera against IyoI, SK05Kyo, Nakajima and Mie0301 isolates were raised in rabbits and used for immobilization assays and Western blotting. Immobilization assays showed that the six isolates could be divided into three groups, tentatively designated serotype I for IyoI, JF05To, RF05To, SK05Kyo, serotype II for Nakajima and serotype III for Mie0301. Western blotting results supported these three serotypes, with marked similarities in the banding profiles of IyoI, JF05To, RF05To and SK05Kyo isolates, which were distinct from the Nakajima and Mie0301 isolates. Three isolates, IyoI, Nakajima and Mie0301 that were selected as representatives of each serotype, were highly pathogenic to Japanese flounder by experimental infection. Based on these findings, we propose that there are at least three M. avidus serotypes in Japan.     

Occurrence of two distinct molecular species of cathepsin B in carp Cyprinus carpio

ABSTRACT:   We purified cathepsins B₁ and B₂ from the ordinary muscle of carp Cyprinus carpio . The N-terminal amino acid sequences (12 residues) of 29 kDa bands of cathepsins B₁ and B₂ are the same and showed high homology of 75% and 83%, respectively, with the heavy chain of rat and human cathepsins B. Based on conserved sequences of other cathepsins B and the N-terminal amino acid sequences of 29 kDa bands, we cloned carp cathepsin B cDNA. The nucleotide sequence of carp cathepsin B cDNA consists of 1470 bp including a 993 bp open reading frame, encoding a deduced protein of 330 amino acids. The deduced amino acid sequence of carp cathepsin B has similarity of 80% to rainbow trout cathepsin B and of 76–78% to other vertebrate cathepsins B. The sequence of its isoform was also determined during molecular cloning, which has 94.8% similarity with first cloned cathepsin B. They are completely same in N-terminal amino acid sequence of heavy chain, active site and potential N-glycosylation site. This indicates there are at least two kinds of cathepsin B functioning in vivo in carp.     

Chemical components and body color of horse mackerel caught in different areas

ABSTRACT: The chemical components and body color of horse mackerel caught between July 1997 and May 2000 were studied. The fish were caught with small- to medium-sized purse seine offshore from Nagasaki and from Tsushima, and with bull trawl seine and medium- to large-sized purse seine in the East China Sea. The crude lipid content of the fish caught offshore from Tsushima was higher than that of the others, and there were no significant differences among the other catches. The extractive nitrogen content of the fish caught with bull trawl seine was somewhat lower than that of the others. The body colors of the fish differed from those suggested by their common names: Kuroaji (black) , Kiaji (yellow), and Shiroaji (white). The crude lipid content and body color indicated that there was no difference in quality among the catches, with the exception of the crude lipid content of the Tsushima catch.