Localized co-transcriptional recruitment of the multifunctional RNA-binding protein CELF1 by lampbrush chromosome transcription units

The highly-extended transcription units of lampbrush chromosomes (LBCs) offer unique opportunities to study the co-transcriptional events occurring on nascent transcripts. Using LBCs from amphibian oocytes, I investigated whether CELF1, an RNA binding protein involved in the regulation of alternative splicing, mRNA stability and translation, is localized to active transcription units. Antibodies raised against mammalian (CUG-BP1) and amphibian (EDEN-BP) CELF1 were used to immunostain LBC spreads prepared from several species, including Xenopus laevis and the axolotl Ambystoma mexicanum. Up to about 50 separate LBC loci were convincingly immunostained and it was clear that CELF1 was present in the nascent RNPs of lateral loops. Furthermore, myc-tagged CUG-BP1 expressed in microinjected axolotl oocytes was specifically targeted to nascent transcripts of loops that recruit endogenous CELF1. In many active transcription units CELF1 was distinctly localized, being first recruited by nascent transcripts only far downstream of the transcription start site and remaining associated until the end of transcription. Overall it appears possible that the multiple functions of CELF1 in regulating posttranscriptional gene expression could all be predetermined during transcription by virtue of a region-specific binding to the nascent transcripts of target genes.