Pax-5 is essential for kappa sterile transcription during Ig kappa chain gene rearrangement.

ABSTRACT

Pax-5 is the key regulator in B cell development. Pax-5-deficient mice show defects in B cell commitment and recombination of IgH chain gene rearrangement from DJ to VDJ. Previously, we found that Pax-5 bound to KI and KII sites, which play a crucial role in kappa-chain gene rearrangement. However, the function of Pax-5 in Ig kappa chain gene rearrangement has not been investigated. To address this issue, we newly established pre-BI cell lines expressing the pre-B cell receptor from Pax-5-deficient mice and used them in an in vitro culture system, in which kappa-chain gene rearrangement is induced by removing IL-7. By examining the Pax-5-deficient pre-BI (knockout (KO)) cells, we show in this study that, despite recombination-activating gene 1 and 2 expression, these KO cells did not rearrange the kappa-chain gene following the absence of kappa sterile transcription. Consistent with these data, fluorescent in situ hybridization analyses revealed that the J(kappa) locus in KO cells was located at the nuclear periphery as a repressive compartment. Transfection of KO cells with Pax-5 constructs indicated that the transactivation domain of Pax-5 was required for kappa sterile transcription and kappa-chain gene rearrangement. Moreover, the hormone-inducible system in KO cells demonstrated that Pax-5 directly functioned in kappa sterile transcription. These results indicate that Pax-5 is necessary for kappa sterile transcription during Ig kappa chain gene rearrangement.