Recombination signal sequences restrict chromosomal V(D)J recombination beyond the 12/23 rule

The genes encoding the variable regions of lymphocyte antigen receptors are assembled from variable (V), diversity (D) and joining (J) gene segments. V(D)J recombination is initiated by the recombinase activating gene (RAG)-1 and -2 proteins, which introduce DNA double-strand breaks between the V, D and J segments and their flanking recombination signal sequences (RSSs). Generally expressed DNA repair proteins then carry out the joining reaction. The conserved heptamer and nonamer sequences of the RSSs are separated by non-conserved spacers of 12 or 23 base pairs (forming 12-RSSs and 23-RSSs). The 12/23 rule, which is mediated at the level of RAG-1/2 recognition and cutting, specifies that V(D)J recombination occurs only between a gene segment flanked by a 12-RSS and one flanked by a 23-RSS. Vbeta segments are appended to DJbeta rearrangements, with little or no direct Vbeta to Jbeta joining, despite 12/23 compatibility of Vbeta 23-RSSs and Jbeta12-RSSs. Here we use embryonic stem cells and mice with a modified T-cell receptor (TCR)beta locus containing only one Dbeta (Dbeta1) gene segment and one Jbeta (Jbeta1) gene cluster to show that the 5' Dbeta1 12-RSS, but not the Jbeta1 12-RSSs, targets rearrangement of a diverse Vbeta repertoire. This targeting is precise and position-independent. This additional restriction on V(D)J recombination has important implications for the regulation of variable region gene assembly and repertoire development.     

Electrophysiological measurement of rapid shifts of attention during visual search.

The perception of natural visual scenes that contain many objects poses computational problems that are absent when objects are perceived in isolation. Vision researchers have captured this attribute of real-world perception in the laboratory by using visual search tasks, in which subjects search for a target object in arrays containing varying numbers of non-target distractor objects. Under many conditions, the amount of time required to detect a visual search target increases as the number of objects in the stimulus array increases, and some investigators have proposed that this reflects the serial application of attention to the individual objects in the array. However, other investigators have argued that this pattern of results may instead be due to limitations in the processing capacity of a parallel processing system that identifies multiple objects concurrently. Here we attempt to address this longstanding controversy by using an electrophysiological marker of the moment-by-moment direction of attention-the N2pc component of the event-related potential waveform--to show that attention shifts rapidly among objects during visual search.