A novel mammalian HORMA domain-containing protein, HORMAD1, preferentially associates with unsynapsed meiotic chromosomes

HORMA domain-containing proteins regulate interactions between homologous chromosomes (homologs) during meiosis in a wide range of eukaryotes. We have identified a mouse HORMA domain-containing protein, HORMAD1, and biochemically and cytologically shown it to be associated with the meiotic chromosome axis. HORMAD1 first accumulates on the chromosomes during the leptotene to zygotene stages of meiotic prophase I. As germ cells progress into the pachytene stage, HORMAD1 disappears from the synapsed chromosomal regions. However, once the chromosomes desynapse during the diplotene stage, HORMAD1 again accumulates on the chromosome axis of the desynapsed homologs. HORMAD1 thus preferentially localizes to unsynapsed or desynapsed chromosomal regions during the prophase I stage of meiosis. Analysis of mutant strains lacking different components of the synaptonemal complex (SC) revealed that establishment of the SC is required for the displacement of HORMAD1 from the chromosome axis. Our results therefore strongly suggest that also mammalian cells use a HORMA domain-containing protein as part of a surveillance system that monitors synapsis or other interactions between homologs.     

Human mitochondrial RNA turnover caught in flagranti: involvement of hSuv3p helicase in RNA surveillance

The mechanism of human mitochondrial RNA turnover and surveillance is still a matter of debate. We have obtained a cellular model for studying the role of hSuv3p helicase in human mitochondria. Expression of a dominant-negative mutant of the hSUV3 gene which encodes a protein with no ATPase or helicase activity results in perturbations of mtRNA metabolism and enables to study the processing and degradation intermediates which otherwise are difficult to detect because of their short half-lives. The hSuv3p activity was found to be necessary in the regulation of stability of mature, properly formed mRNAs and for removal of the noncoding processing intermediates transcribed from both H and L-strands, including mirror RNAs which represent antisense RNAs transcribed from the opposite DNA strand. Lack of hSuv3p function also resulted in accumulation of aberrant RNA species, molecules with extended poly(A) tails and degradation intermediates truncated predominantly at their 3′-ends. Moreover, we present data indicating that hSuv3p co-purifies with PNPase; this may suggest participation of both proteins in mtRNA metabolism.     

Crystallization of alcohol oxidase fromPichia pastoris. Secondary structure predictions indicate a domain with the eightfold β/α-barrel fold

Alcohol oxidase fromPichia pastoris has been crystallized from polyethylene glycol 4000 solutions. The crystals are tetragonal, a=228 Å, c=456 Å space groupP4₁2₁2. The crystals scatter only to about 6 Å resolution; their poor crystallinity may have some physiological function. Secondary structure predictions suggest that the C-terminal part of the molecule, residues 311–664, has the folding of an eightfold /-barrel (TIM barrel). This would indicate common ancestry with four other flavoenzymes: canavalin, glycolate oxidase, flavocytochrome b, and trimethylamine dehydrogenase.     

Reduced insecticide susceptibility among populations of Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) in strawberry production

Chilli thrips, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) is a major, economically important, and recent invasive pest of strawberries and other horticultural crops in United States. Several conventional insecticides are used for S. dorsalis management, and resistance development threatens loss of few available tools. Hence, our objectives were to: (1) determine the susceptibility of S. dorsalis to commonly used conventional insecticides: spinetoram, acetamiprid, cyantraniliprole and bifenthrin, and (2) establish LC₅₀ and LC₉₀ dosages for spinetoram against S. dorsalis. Sampling of S. dorsalis populations was conducted twice in seven strawberry fields in Florida during the strawberry field season between 2019 and 2020. Leaf-disc bioassays were performed with field collected populations along with a susceptible 2-year-old laboratory culture of S. dorsalis. Overall, at highest recommended rate the percent mortality of late season S. dorsalis populations from five out of seven collection sites was lower (~41%) than average mortality observed with early season populations (~72%). Populations from at least four out of seven sampling sites exhibited significantly lower mortality than the laboratory susceptible culture in late season. The LC₅₀ and LC₉₀ values for spinetoram for the susceptible laboratory population were 0.026 and 8.64 ppm, respectively. On the other hand, LC₅₀ values of field collected populations to spinetoram varied with resistance ratios ranging from 6 to 269 fold as compared against the laboratory strain. Our results suggest that susceptibility of S. dorsalis to commonly used insecticides in strawberries varies significantly between early and late season populations within the same crop season. The efficacy of bifenthrin against S. dorsalis was particularly low (~ reduced to half in late season), especially among field collected populations. Our results indicate an urgent need to incorporate other pest management strategies, as well as effective rotation programs to reduce selection for resistance among populations of S. dorsalis in strawberry production.     

Adaptation of anaerobically grown Thauera aromatica,Geobacter sulfurreducens and Desulfococcus multivorans to organic solvents on the level of membrane fatty acid composition

The effect of different solvents and pollutants on the cellular fatty acid composition of three bacterial strains: Thauera aromatica, Geobacter sulfurreducens and Desulfococcus multivorans, representatives of diverse predominant anaerobic metabolisms was investigated. As the prevailing adaptive mechanism in cells of T. aromatica and G. sulfurreducens whose cellular fatty acids patterns were dominated by palmitic acid (C16:0) and palmitoleic acid (C16:1cis), the cells reacted by an increase in the degree of saturation of their membrane fatty acids when grown in the presence of sublethal concentrations of the chemicals. Next to palmitic acid C16:0, the fatty acid pattern of D. multivorans was dominated by anteiso-branched fatty acids which are characteristic for several sulfate-reducing bacteria. The cells responded to the solvents with an increase in the ratio of straight-chain saturated (C14:0, C16:0, C18:0) to anteiso-branched fatty acids (C15:0anteiso, C17:0anteiso, C17:1anteisoΔ9cis). The results show that anaerobic bacteria react with similar mechanisms like aerobic bacteria in order to adapt their membrane to toxic organic solvents. The observed adaptive modifications on the level of membrane fatty acid composition can only be carried out with de novo synthesis of the fatty acids which is strictly related to cell growth. As the growth rates of anaerobic bacteria are generally much lower than in the so far investigated aerobic bacteria, this adaptive response needs more time in anaerobic bacteria. This might be one explanation for the previously observed higher sensitivity of anaerobic bacteria when compared with aerobic ones.     

Behavioral evidence for a female-produced sex attractant in Diaphorina citri

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important worldwide pest of citrus. It vectors three phloem-restricted bacteria in the genus Candidatus Liberibacter that cause huanglongbing (citrus greening disease). Studies were conducted to examine the behavioral responses of male and female D. citri to conspecifics of the same and opposite sex, with and without associated citrus host plants, in both open-air arena choice assays and Y-tube olfactometer assays. Virgin and mated male D. citri colonized citrus plants that were currently or had been previously colonized by virgin or mated female D. citri in greater numbers than control plants without females. However, males or females did not accumulate more on plants colonized by conspecifics of the same sex compared to uninfested plants, and females showed no preference for plants pre-infested with males compared with uninfested controls. In complementary Y-tube olfactometer assays, virgin and mated males chose arms with odor sources from mated females compared with blank controls in the absence of associated citrus host plant volatiles. In both behavioral assays, mated female D. citri appeared more attractive than virgin females. The vibrational calling behavior of male D. citri was reduced when males were challenged by the odors of conspecific mated females relative to when males were challenged by the odor of other males. Collectively, our results provide behavioral evidence for a female-produced volatile sex attractant pheromone in D. citri . Future identification and synthesis of a sex attractant pheromone will be an important contribution to current monitoring and management practices for D. citri .     

Molecular basis of the dual functions of 2B4 (CD244)

2B4 belongs to the CD2 family of molecules and is expressed on all NK, gammadelta, and memory CD8(+) (alphabeta) T cells. The murine NK receptor 2B4 exhibits both inhibitory and activating functions, whereas human 2B4 has been reported to be an activating molecule. How murine 2B4 can act both as an activating and inhibitory receptor and what distinguishes its function from human 2B4 have remained largely unknown. We use here a model system that allows the study of human and murine 2B4 under identical and controlled conditions. These studies reveal that both human and mouse 2B4 can activate or inhibit NK cells. We show here that the level of 2B4 expression and the degree of 2B4 cross-linking play a significant role in the regulation of signaling lymphocyte activation molecule-associated protein-mediated activation by 2B4. A high level of 2B4 expression, heavy cross-linking, and relative paucity of signaling lymphocyte activation molecule-associated protein promote inhibitory function. Our studies demonstrate how a single receptor can have opposing function depending on the degree of receptor expression, extent of its ligation, and the relative abundance of certain adaptor molecules. Because the levels of 2B4 and CD48 are dynamically regulated, these findings have implications for the regulation of NK cell function.