Serino G, Su H, Peng Z, Tsuge T, Wei N, Deng XW (2003) Characterization of the last subunit of the arabidopsis COP9 signalosome: Implications for the overall structure and origin of the complex. Plant Cell 15(3): 719-731.
The COP9 signalosome (CSN) is an evolutionarily conserved protein complex that resembles the lid subcomplex of proteasomes. Through its ability to regulate specific proteasome-mediated protein degradation events, CSN controls multiple aspects of development. Here, we report the cloning and characterization of AtCSN2, the last uncharacterized CSN subunit from Arabidopsis. We show that the AtCSN2 gene corresponds to the previously identified FUS12 locus and that AtCSN2 copurifies with CSN, confirming that AtCSN2 is an integral component of CSN. AtCSN2 is not only able to interact with the SCF(TIR1) subunit AtCUL1, which is partially responsible for the regulatory interaction between CSN and SCF(TIR1), but also interacts with AtCUL3, suggesting that CSN is able to regulate the activity of other cullin-based E3 ligases through conserved interactions. Phylogenetic analysis indicated that the duplication and subsequent divergence events that led to the genes that encode CSN and lid subunits occurred before the divergence of unicellular and multicellular eukaryotic organisms and that the CSN subunits were more conserved than the lid subunits during evolution. Comparative analyses of the subunit interaction of CSN revealed a set of conserved subunit contacts and resulted in a model of CSN subunit topology, some aspects of which were substantiated by in vivo cross-link tests.
Peterson DG, Wessler SR, Paterson AH (2002) Efficient capture of unique sequences from eukaryotic genomes. Trends in Genetics 18(11): 547-550.
Cot-based cloning and sequencing (CBCS), a synthesis of Cot analysis, DNA cloning and high-throughput sequencing, promises to accelerate the study of eukaryotic genomes. In particular, CBCS will (1) permit efficient gene discovery in species with substantial quantities of repetitive DNA, (2) allow the sequence complexity (i.e. all the unique sequence information) of large genomes to be elucidated at a fraction of the cost of shotgun sequencing, and (3) enhance genome sequencing efforts by facilitating capture of low-copy sequences not secured by EST sequencing. CBCS should accelerate comparative genomics research, especially in large genomes such as those of many crops.
Pechan T, Cohen A, Williams WP, Luthe DS (2002) Insect feeding mobilizes a unique plant defense protease that disrupts the peritrophic matrix of caterpillars. Proceedings of the National Academy of Sciences of the United States of America 99(20): 13319-13323.
Plants frequently respond to herbivorous insect attack by synthesizing defense proteins that deter insect feeding and prevent additional herbivory. Maize (Zea mays L.) lines, resistant to feeding by a number of lepidopteran species, rapidly mobilize a unique 33-kDa cysteine protease in response to caterpillar feeding. The accumulation of the 33-kDa cysteine protease in the maize mid-whorl was correlated with a significant reduction in caterpillar growth that resulted from impaired nutrient utilization. Black Mexican Sweetcorn callus transformed with mir1, the gene encoding the 33-kDa cysteine protease, expressed the protease and growth of caterpillars reared on the transgenic callus was reduced 60-80%. Scanning electron microscopy was used to examine the effect of plant material expressing the 33-kDa cysteine protease on the structure of the caterpillar peritrophic matrix. Because the peritrophic matrix surrounds the food bolus, assists in digestive processes, and protects the caterpillar midgut from physical and chemical damage, disruption of peritrophic matrix may reduce caterpillar growth. The results indicated that the peritrophic matrix was severely damaged when caterpillars fed on resistant maize plants or transgenic Black Mexican Sweetcorn. The accumulation of the 33-kDa cysteine protease in response to caterpillar feeding, and its ability to damage the insect peritrophic matrix, represents an unusual host-plant resistance mechanism that may have applications in agricultural biotechnology.
Burnham MR, Branton SL, Peebles ED, Lott BD, Gerard PD (2002) Effects of F-strain Mycoplasma gallisepticum inoculation at twelve weeks of age on performance and egg characteristics of commercial egg-laying hens. Poultry Science 81(10): 1478-1485.
The effects of F-strain Mycoplasma gallisepticum (FMG) inoculation during the pullet period on the subsequent performance and egg characteristics of commercial Single Combed White Leghorn hens were evaluated. In two trials, BW, feed consumption, egg production (EP), egg weight, egg size class, relative eggshell water vapor conductance, and relative percentages of eggshell, yolk and albumen weights were determined through approximately 60 wk of age. In each trial, pullets at 12 wk of age were randomly assigned to negative pressure biological isolation units. Birds in one-half of the total units were inoculated with FMG, and the other half were sham-inoculated with sterile media. In both trials, onset of lay was delayed approximately 1 wk in layers inoculated with FMG. Control birds that had not been previously inoculated with FMG laid their first egg at 18 wk of age, while birds that had been previously inoculated with FMG laid their first egg at 19 wk of age. In Trial 1, FMG-inoculated hens laid significantly fewer total eggs, which became apparent at each week after Week 42. In Trial 2, a numerical decrease in total EP occurred, and the percentage of undersized eggs laid by FMG-inoculated birds was significantly lower at 19 wk of age but was higher at 20 and 21 wk when compared to controls. Mortality was not significantly different between the treatments in either trial. These data demonstrate that when birds are housed in isolation facilities and inoculated with FMG at 12 wk of age, onset of lay is delayed. These data also suggest that FMG may lead to delays in undersize EP and decreases in total EP. However, because significant FMG effects on these parameters were observed in only one trial, additional studies may be necessary to verify these effects.