The chicken Ig light chain 3'-enhancer is essential for gene expression and regulates gene conversion via the transcription factor E2A.
Expression of the rearranged chicken immunoglobulin light chain (IgL) gene is regulated by a V gene promoter, a matrix attachment region (MAR) in the J-C intron and an enhancer downstream of the Ig constant region. Using knockout analysis, we demonstrate that the 3'-enhancer is not only required for gene activation but is also essential for the maintenance of gene expression. Deletion of the MAR on the other hand increases IgL transcription, indicating that the MAR acts as negative regulator. We demonstrate that Id1 and Id3, dominant-negative regulators of basic-region helix-loop-helix (bHLH) transcription factors, are able to reduce chicken IgL 3'-enhancer activity in transient assays and strongly reduce the rate of gene conversion (GC) in DT40 clone 18 cells. Conversely, overexpression of avian E47, a bHLH transcription factor, leads to a dramatic increase in GC rates independent of IgL or activation-induced cytidine deaminase RNA levels. Thus, E47 is the first transcription factor to activate GC without an apparent increase in transcription.
Conlon
Thomas M
TM
Cambridge Institute for Medical Research and Department of Pathology, Cambridge University, Cambridge, UK.
Meyer
Kerstin B
KB
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D002645:Chickens / Q000276:immunology*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005434:Flow Cytometry; D005785:Gene Conversion; D015870:Gene Expression; D005786:Gene Expression Regulation; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D045170:Matrix Attachment Regions / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015533:Transcriptional Activation
D051792:Basic Helix-Loop-Helix Proteins; D007147:Immunoglobulin Light Chains