An amphitropic cAMP-binding protein in yeast mitochondria

ABSTRACT: We describe the first example of a mitochondrial protein with a covalently attached phos-phatidylinositol moiety acting as a membrane anchor. The protein can be metabolically labeled with both stearic acid and inositol. The stearic acid label is removed by phospholipase D whereupon the protein with the retained inositol label is released from the membrane. This protein is a cAMP receptor of the yeast Saccharomyces cereuisiae and tightly associated with the inner mitochondrial membrane. However, it is converted into a soluble form during incubation of isolated mitochondria with Ca2+ and phospholipid (or lipid derivatives). This transition requires the action of a proteinaceous, N-ethylmaleimide-sensitive component of the intermembrane space and is accompanied by a decrease in the lipophilicity of the cAMP receptor. We propose that the component of the intermembrane space triggers the amphitropic behavior of the mitochondrial lipid-modified CAMP-binding protein through a phospholipase activity. Only in recent years specific fatty acids have been recog-nized to play important roles in the association of proteins with membranes. Both noncovalent and covalent interactions be-tween fatty acids and proteins have been reported. Among the latter are GTP-binding proteins (Molenaar et al., 1988)

Identification and characterization of Rhipicephalus (Boophilus) microplus candidate protective antigens for the control of cattle tick infestations

The cattle ticks, Rhipicephalus (Boophilus) spp., affect cattle production in tropical and subtropical regions of the world. Tick vaccines constitute a cost-effective and environmentally friendly alternative to tick control. The recombinant Rhipicephalus microplus Bm86 antigen has been shown to protect cattle against tick infestations. However, variable efficacy of Bm86-based vaccines against geographic tick strains has encouraged the research for additional tick-protective antigens. Herein, we describe the analysis of R. microplus glutathione-S transferase, ubiquitin (UBQ), selenoprotein W, elongation factor-1 alpha, and subolesin (SUB) complementary DNAs (cDNAs) by RNA interference (RNAi) in R. microplus and Rhipicephalus annulatus. Candidate protective antigens were selected for vaccination experiments based on the effect of gene knockdown on tick mortality, feeding, and fertility. Two cDNA clones encoding for UBQ and SUB were used for cattle vaccination and infestation with R. microplus and R. annulatus. Control groups were immunized with recombinant Bm86 or adjuvant/saline. The highest vaccine efficacy for the control of tick infestations was obtained for Bm86. Although with low immunogenic response, the results with the SUB vaccine encourage further investigations on the use of recombinant subolesin alone or in combination with other antigens for the control of cattle tick infestations. The UBQ peptide showed low immunogenicity, and the results of the vaccination trial were inconclusive to assess the protective efficacy of this antigen. These experiments showed that RNAi could be used for the selection of candidate tick-protective antigens. However, vaccination trials are necessary to evaluate the effect of recombinant antigens in the control of tick infestations, a process that requires efficient recombinant protein production and formulation systems

A novel species of double stranded RNA in mitochondria of Saccharomyces cerevisiae.

A double stranded RNA species has been detected in guanidine hydrochloride extracts of mitochondria from respiratory competent cells of Saccharomyces cerevisiae. This novel mitochondrial RNA, termed mtdsRNA, has been purified in a Cs2SO4 density gradient where it bands at a density of 1.58 g/ml. The mtdsRNA runs as a single slow moving band on agarose gels. Its double stranded RNA character was evidenced by its sensitivity to digestion by RNase III, but not by RNase H, or DNase I. Moreover the mtdsRNA hybridized to each separated strand of a petite mtDNA. It is concluded that mtdsRNA contains long transcripts derived from most regions of yeast mtDNA, because 1) its weight-average length as determined by electron microscopy was 4.5 micrometer (about 14 kb, or 20% of the wild type mtDNA genome), and 2) it hybridized to each of a series of eight petite mtDNA probes carrying sequences derived from widely different segments of mtDNA. It is proposed that prolonged transcription of both strands of yeast mtDNA can occur and that mtdsRNA arises from hybridization of these long complementary transcripts

Successful vaccination against Boophilus microplus and Babesia bovis using recombinat antigens

Current methods for the control of the cattle tick Boophils microplus and the agent of bovine babesiosis, Babesia bovis are unsatisfactory. Effective immunological control of both parasites would have great advantages. However, naturally acquired immunity to the tick is generally unable to prevent serious production losses. A vaccine against the tick, based on a novel form of immunization, is being developed. A protective antigen has been isolated from the tick, characterized and produced as an effective, recombinant protein. A vaccine incorporating this antigen is currently undergoing field trials. In the Australian situation, improved tick control will probably increase endemic instability with respect to B. bovis. Fortunately, a trivalent, recombinant B. bovis vaccine has also been developed. This too is now undergoing pre-registration field trials

Queensland’s multi-year Wet and Dry periods: implications for grazing enterprises and pasture resources

Year-to-year variability in rainfall has long been recognised as a major issue in managing livestock enterprises across Australia's grazing lands. Extension products documenting rainfall variability have been developed over the last 30 years and have been keenly sought by producers and their advisors. This paper describes multi-year rainfall variability from 1889 to 2020 and provides the basis for classifying the 131 years of rainfall into 18 discrete Wet (7), Average (2) and Dry (9) periods as presented in the 'Queensland's Extended Wet and Dry Periods' poster. The classification was consistent with: Analysis of fluctuations and trends in the long-term time series of reported livestock numbers; drought declarations for government assistance; and documented periods of pasture resource degradation and recovery. Rainfall during the nine Wet and Average periods was +18% above the long-term average annual rainfall (LTAAR), in contrast to the Dry periods with-17% below LTAAR. Wet periods (including Average) were on average 7 years in duration, ranging from 5 to 9 years. Dry periods were on average 8 years in duration and ranged from 5 to 13 years. Detailed analysis of the effects of the El Nino Southern Oscillation (ENSO) phenomenon indicated that: (a) the Wet/Dry periods were dominated by different frequencies and amounts of rainfall in La Nina/El Nino years; (b) rainfall in ENSO neutral years was generally above and below average rainfall for the Wet or Dry periods respectively; (c) the frequency of ENSO year-types was less important than the overall rainfall surplus (or deficit) in La Nina (or El Nino) years within the Wet (or Dry) periods respectively; and (d) the timing of Wet and Dry periods was correlated with indices of quasi-decadal and inter-decadal variability in components (sea surface temperatures and atmospheric pressures) of the global climate system. Climatic risk assessment systems for grazing management at multi-year timescales are yet to be developed

Potential Climate Change Impacts on Beef Production Systems in Australia

There is increasing evidence suggesting that Australia\u27s climate is changing due to enhanced levels of greenhouse gases and that it will continue to change (Pittock 2003). Climate changes are partly established, however the impact on systems, industries and process are unclear. Industry distribution reflects climatically imposed boundaries and the relative profitability of alternative land use. Climate change may negatively impact some existing industries but create opportunities for others. This study provides an assessment of the likely impacts of plausible climate change on the beef industry in central Queensland