A CD45 polymorphism associated with abnormal splicing is absent in African populations

The CD45 antigen is essential for normal antigen receptor-mediated signalling in lymphocytes, and different patterns of splicing of CD45 are associated with distinct functions in lymphocytes. Abnormal CD45 splicing has been recognized in humans, caused by a C77G transversion in the gene encoding CD45 (PTPRC). Recently the C77G polymorphism has been associated with multiple sclerosis and increased susceptibility to HIV-1 infection. These studies suggest that the regulation of CD45 splicing may be critical for the proper function of the immune system. Because of these data we examined the frequency of the C77G allele in African and Asian populations from countries with high or low prevalence of HIV infection. Here we report that the variant CD45 C77G allele is absent in African populations. We further show that populations living in the Pamir mountains of Central Asia have a very high prevalence of the C77G variant.     

Does Mangrove Leaf Chemistry Help Explain Crab Herbivory Patterns?

We examined feeding by the mangrove tree crab Aratus pisonii in Tampa Bay, Florida, in relation to the percent dry weight of carbohydrate, protein, phenolics, condensed tannins, ash, carbon, nitrogen, carbonmitrogen ratio, water content, and sclerophylly for leaves of the red mangrove Rhizophora mangle. Comparisons of leaf chemistry were made among leaves that experienced variable levels of crab damage. Because R. mangle is the crab's preferred food source based on damage patterns in the field, comparisons of R. mangle leaf chemistry were made in relation to that of the black mangrove Avicennia germinans and the white mangrove Laguncularia racemosa. We observed a negative relationship between level of leaf damage and percent dry weight of nitrogen, carbohydrates, condensed tannins, and sclerophylly. In contrast, a positive relationship was found between leaf damage and the carbon:nitrogen ratio. The chemical constituents that provided the best explanation for differences in damage among the three mangrove species include condensed tannins, nitrogen, carbon:nitrogen ratio, carbohydrates, phenolics, water content, and ash. The results from this study suggest that chemistry only partially explains food preference by A. pisonii. It appears that A. pisonii feeding behavior and preference may be influenced by a more complex series of factors and interactions, which may include reproduction by, predation on, and interspecific competition with A. pisonii.     

The Yeast Saccharomyces cerevisiae Contains Two Glutaredoxin Genes That Are Required for Protection against Reactive Oxygen Species

Glutaredoxins are small heat-stable proteins that act as glutathione-dependent disulfide oxidoreductases. Two genes, designated GRX1 and GRX2, which share 40–52% identity and 61–76% similarity with glutaredoxins from bacterial and mammalian species, were identified in the yeast Saccharomyces cerevisiae. Strains deleted for both GRX1 and GRX2 were viable but lacked heat-stable oxidoreductase activity using β-hydroxyethylene disulfide as a substrate. Surprisingly, despite the high degree of homology between Grx1 and Grx2 (64% identity), the grx1 mutant was unaffected in oxidoreductase activity, whereas the grx2 mutant displayed only 20% of the wild-type activity, indicating that Grx2 accounted for the majority of this activity in vivo. Expression analysis indicated that this difference in activity did not arise as a result of differential expression of GRX1 and GRX2. In addition, a grx1 mutant was sensitive to oxidative stress induced by the superoxide anion, whereas a strain that lacked GRX2 was sensitive to hydrogen peroxide. Sensitivity to oxidative stress was not attributable to altered glutathione metabolism or cellular redox state, which did not vary between these strains. The expression of both genes was similarly elevated under various stress conditions, including oxidative, osmotic, heat, and stationary phase growth. Thus, Grx1 and Grx2 function differently in the cell, and we suggest that glutaredoxins may act as one of the primary defenses against mixed disulfides formed following oxidative damage to proteins.     

Fate and role of peroxisomes during the life cycle of the yeast Saccharomyces cerevisiae: inheritance of peroxisomes during meiosis

 Sporulation in the yeast Saccharomyces cerevisiae is a meiotic developmental process that occurs in MAT a/MATα heterozygotes in response to nutrient deprivation. Here, the fate and role of peroxisomes during sporulation and germination has been examined by a combination of immunoelectron microscopy and the use of pex mutants defective in peroxisomal functions. Using a green fluorescent protein probe targeted to peroxisomes we show that peroxisomes are inherited through meiosis and that they do not increase in number either during sporulation or spore germination. In addition, there is no requirement for peroxisome degradation prior to spore packaging. Unlike the situation in filamentous fungi, peroxisomes do not proliferate during the yeast life cycle. Functional peroxisomes are dispensable for efficient meiotic development on acetate medium since homozygous Δpex6 diploids sporulated well and produced mature spores that were resistant to diethyl ether. Like haploids, diploid cells can proliferate their peroxisomes in response to oleate as sole carbon source in liquid medium, but under these conditions they do not sporulate. On solid oleate medium, homozygous pex5,Δpex6, and pex7 cells were unable to sporulate efficiently, whereas the wild type was. The results presented here are discussed in terms of the transmission of organelles to progeny cells. Accepted: 19 December 1997     

Temperature-sensitive Saccharomyces cerevisiae mutant defective in lipid biosynthesis.

A temperature-sensitive mutant of Saccharomyces cerevisiae (DAM303) is described that exhibits an early defect in lipid biosynthesis at the restrictive growth temperature, 37 degrees C. This strain rapidly lost viability after 1 h of incubation at 37 degrees C, and this was accompanied by a significantly reduced incorporation of 32Pi into cellular lipid and an accumulation of [1-14C]acetate into the free fatty acid fraction. The temperature-sensitive DAM303 mutation failed to complement the sec13 mutation described by Novick et al. (Cell 21:205-215, 1980), and from analysis of invertase secretion in the temperature-sensitive DAM303 strain, it is clear that the loss of invertase secretion in the mutant occurs after the loss of phospholipid synthesis. Although the precise nature of the temperature-sensitive lesion in the DAM303 strain has still to be identified, the results from the study of this mutant indicate that a defect in lipid biosynthesis can be correlated with subsequent alterations in extracellular protein secretion and loss of other macromolecular functions including DNA, RNA, and protein syntheses. From studies of this mutant, two procedures of enriching for other temperature-sensitive mutants with defects in lipid biosynthesis have emerged: inositol overproduction and screening for increased buoyant densities.     

Optimal conditions for selecting specific auxotrophs of Saccharomyces cerevisiae using temperature-sensitive suicide mutants.

Of 48 temperature-sensitive mutants of Saccharomyces cerevisiae examined, five belonging to the same complementation group were found to undergo extensive loss of viability at the restrictive temperature. These mutants were protected from the lethal effects of exposure to a non-permissive temperature by starving for an auxotrophic requirement. By analogy with the method described by Littlewood [6] for selecting antibiotic-sensitive mutants, these temperature-sensitive mutants were found suitable in enriching for specific auxotrophs. Optimal conditions have been determined for selecting specific auxotrophs after mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine. These enable 20-fold enrichment and at least in the case of mutation to adenine dependence the method does not appear to favour mutations at any particular locus.     

Restriction endonuclease analysis for the identification of baculovirus pesticides.

Gel electrophoresis of deoxyribonucleic acid (DNA) fragments generated by digesting the DNA genomes of nuclear polyhedrosis viruses (NPV) with restriction endonucleases provides DNA fragment patterns that may be used to identify different viruses of this group. Characteristic fragment patterns were obtained for three NPVs, which are important as biological pesticides (Autographa californica NPV, Orgyia pseudotsugata NPV, and Heliothis zea NPV). The DNA fragment patterns of the A. californica NPV genoms did not change with passage through the alternate insect host, Trichoplusia ni. Heterogeneity in one preparation of O. pseudotsugata NPV was observed. The identification procedure is direct and precise. Applications of this procedure include quality control of commercial preparations of viral pesticides and screening for genetic alterations in the viruses.