Characterization of the Arabidopsis lecRK-a genes: members of a superfamily encoding putative receptors with an extracellular domain homologous to legume lectins.

An Arabidopsis cDNA clone that defines a new class of plant serine/threonine receptor kinases was found to be a member of a family of four clustered genes (lecRK-a1-a4) which have been cloned, sequenced and mapped on chromosome 3. This family belongs to a large superfamily encoding putative receptors with an extracellular domain homologous to legume lectins and appears to be conserved at least among dicots. In the Columbia ecotype only the lecRK-a1 and perhaps the lecRK-a3 gene is functional, since lecRK-a2 is disrupted by a Ty-copia retroelement and lecRK-a4 contains a frameshift mutation. Structural analysis of the lecRK-al and lecRK-a3 deduced amino-acid sequences suggests that the lectin domain is unlikely to be involved in binding monosaccharides but could interact with complex glycans and/or with hydrophobic ligands. Immunodetection of lecRK gene products in plasma membranes purified by free-flow electrophoresis showed that the lecRK-a proteins are probably highly glycosylated integral plasma membrane components.

Genome DNA sequencing around the EF-1 alpha multigene locus of Arabidopsis thaliana indicates a high gene density and a shuffling of noncoding regions.

In Arabidopsis thaliana, EF-1 alpha proteins are encoded by a multigene family of four members. Three of them are clustered at the same locus, which was positioned 24 cM from the top of chromosome 1. A region of DNA spanning 63 kb around these locus was sequenced and analyzed. One main characteristic of the locus is the mosaic organization of both genes and intergenic regions. Fourteen genes were identified, among which only four were already described, and other unidentified are most likely present. Functionally diverse genes are found at close intervals. Exon and intron distribution is highly variable at this locus, one gene being split into at least 20 introns. Several duplications were found within the sequenced segment both in coding and noncoding regions, including two gene families. Moreover, a sequence corresponding to the 5' noncoding region of the EF-1 alpha genes and harboring a 5' intervening sequence is duplicated and found upstream of several genes, suggesting that noncoding regions can be shuffled during evolution.

Characterization of an Arabidopsis thaliana gene that defines a new class of putative plant receptor kinases with an extracellular lectin-like domain.

We have characterized an Arabidopsis receptor-like serine/threonine kinase gene, Ath.lecRK1 (Arabidopsis thaliana lectin-receptor kinase), defining a new and putatively important class of plant receptor kinases. Structural features of the predicted polypeptide include an amino-terminal membrane-targeting signal sequence, a legume lectin-like extracellular domain, a single membrane-spanning domain, and a characteristic serine/threonine protein kinase domain. A recombinant protein containing the kinase domain can be autophosphorylated on a serine residue. Ath.lecRK1 is a member of a gene family of at least two closely related genes. Northern blot analysis indicates that the Ath.lecRK1 gene is weakly expressed in a variety of organs and is regulated in Arabidopsis cell suspension cultures according to the growth phase of cells. The role this new class of plant receptor kinase could play is discussed with regard to the transduction of oligosaccharide and plant hormone signals.

Noninvasive external recording of cardiac conduction system (His bundle) activity.

Successful and adequate external recording of the cardiac conduction system from the body's surface can be accomplished in 80 to 90 percent of subjects studied. High-gain amplification, signal averaging, and triggering with a conditioned QRS signal results in good recording reproducibility. Averaging of 128 consecutive cycles is adequate, but on occasion averaging of 256 cycles may yield better results. The patients's QRS signal triggers the transfer of signals, which are digitized and stored during the preceding P-R interval. Comparison of external recordings with direct invasive recordings in animals and patients shows good correlation between the major His bundle deflections. The advantages of the system developed include its mobility, triggering the QRS with pretrigger data processing, and instantaneous display on Polaroid photograph. Future research should concentrate on further miniaturization and simplification of the instrumentation, detailed experimental comparison between direct and external recordings for identification of deflections and their origin, further study of the recording lead system, and the most appropriate method of information display.

Noninvasive recording of His-Purkinje activity in man by ORS-triggered signal averaging.

Mobile instrumentation and a clinically applicable method have been developed for external His bundle recording. High gain signal amplification (10)(5) filtering (30--300 HZ) and averaging (128 or 256 consecutive cycles) are used. Acquisition of signals arising in the P-R interval is triggered by the patient's QRS signal at the end of that interval. The precordial bipolar electrodiogram is digitized at 5k HZ with 8 bit resolution and transferred to a 1,024 word, 18 bit signal averager. The averaged signal is then displayed on an oscilloscope and photographed. Good correlations were obtained between direct intracardiac and precordial recordings in experimental animals and in humans. Noise level after averaging was below 0.3 microV, and there was good elimination of asynchronous atrial and ectopic ventricular activity. With averaging of 128 or 256 consecutive cycles, the signal attenuation after propagation to the chest wall was in the range 1:2000 to 1:4000 in comparison with the directly recorded His bundle activity deflections. The noninvasive method may be of value in follow-up of acute and chronic disturbances of atrioventricular conduction, as well as in studies of effects of pharmacologic interventions.