Identification of two topologically independent domains in RAG1 and their role in macromolecular interactions relevant to V(D)J recombination.

V(D)J recombination is instigated by the recombination-activating proteins RAG1 and RAG2, which catalyze site-specific DNA cleavage at the border of the recombination signal sequence (RSS). Although both proteins are required for activity, core RAG1 (the catalytically active region containing residues 384-1008 of 1040) alone displays binding specificity for the conserved heptamer and nonamer sequences of the RSS. The nonamer-binding region lies near the N terminus of core RAG1, whereas the heptamer-binding region has not been identified. Here, potential domains within core RAG1 were identified using limited proteolysis studies. An iterative procedure of DNA cloning, protein expression, and characterization revealed the presence of two topologically independent domains within core RAG1, referred to as the central domain (residues 528-760) and the C-terminal domain (residues 761-980). The domains do not include the nonamer-binding region but rather largely span the remaining relatively uncharacterized region of core RAG1. Characterization of macromolecular interactions revealed that the central domain bound to the RSS with specificity for the heptamer and contained the predominant binding site for RAG2. The C-terminal domain bound DNA cooperatively but did not show specificity for either conserved RSS element. This domain was also found to self-associate, implicating it as a dimerization domain within RAG1.

Molecular contingencies in schedules of intermittent punishment.

In two experiments, key pecking of pigeons was maintained by a variable-interval 180-s schedule of food presentation. Conjointly, a second schedule delivered response-dependent electric shock. In the first experiment, shocks were presented according to either a variable-interval or a nondifferential interval-percentile schedule. The variable-interval shock schedule differentially delivered shocks following long interresponse times. Although the nondifferential shock schedules delivered shocks less differentially with respect to interresponse times, the two shock schedules equally reduced the relative frequency of long interresponse times. The second experiment differentially shocked long or short interresponse times in different conditions, with resulting decreases in the relative frequency of the targeted interresponse times. These experiments highlight the importance of selecting the appropriate level of analysis for the interaction of behavior and environment. Orderly relations present at one level of analysis (e.g., interresponse times) may not be revealed at other levels of analysis (e.g., overall response rate).

Changes in functional response units with briefly delayed reinforcement.

In two experiments, key-peck responding of pigeons was compared under variable-interval schedules that arranged immediate reinforcement and ones that arranged unsignaled delays of reinforcement. Responses during the nominal unsignaled delay periods had no effect on the reinforcer presentations. In Experiment 1, the unsignaled delays were studied using variable-interval schedules as baselines. Relative to the immediate reinforcement condition, 0.5-s unsignaled delays decreased the duration of the reinforced interresponse times and increased the overall frequency of short (<0.5-s) interresponse times. Longer, 5.0-s unsignaled delays increased the duration of the reinforced interresponse times and decreased the overall frequency of the short interresponse times. In Experiment 2, similar effects to those of Experiment 1 were obtained when the 0.5-s unsignaled delays were imposed upon a baseline schedule that explicitly arranged reinforcement of short interresponse times and therefore already generated a large number of short interresponse times. The results support earlier suggestions that the unsignaled 0.5-s delays change the functional response unit from a single key peck to a multiple key-peck unit. These findings are discussed in terms of the mechanisms by which contingencies control response structure in the absence of specific structural requirements.

A role for negative reinforcement of response omission in punishment?

This experiment attempted to disentangle response-rate reductions controlled by the direct suppressive effects of a punisher from those due to negative reinforcement of response omission. Key-peck responding of pigeons was maintained by a conjoint variable-interval 3-min schedule of food presentation variable-interval 30-s schedule of response-dependent electric shock presentation. Omission of responses for 5, 10, or 30 s resulted in the possibility of canceling a scheduled shock. Response rates were a function of required pause duration, with lower rates occurring when longer periods of response omission were required for shock cancellation. These results show that, with several parameters of punishment held constant, response rates were controlled by the negative reinforcement contingency. Such a finding argues for renewed consideration of the role of negative reinforcement in punishment contingencies.