Chromosomal storage of the RNA-editing enzyme ADAR1 in Xenopus oocytes.
ADARs (adenosine deaminases that act on RNA) are RNA-editing enzymes that convert adenosines to inosines in structured or double-stranded RNAs. Expression and intracellular distribution of ADAR1 is controlled by a plethora of mechanisms suggesting that enzyme activity has to be tightly regulated. Mammalian ADAR1 is a shuttling protein, whereas Xenopus ADAR1 is exclusively nuclear. In oocytes, Xenopus ADAR1 associates with most nascent transcripts but is strongly enriched at a specific site on chromosome 3, termed the special loop. Enrichment at this site requires the presence of RNAs but is independent of ongoing transcription. Here we show that RNAs transcribed elsewhere in the genome accumulate at the special loop even in the absence of transcription. In situ hybridization experiments, however, indicate the absence of known editing substrates from this site. In the absence of transcription also other RNA binding and processing factors accumulate at the special loop, suggesting that ADAR1 is stored or assembled at the special loop in an RNA-containing complex. Nuclear injection of RNAs providing binding sites for ADAR1 dissociates the enzyme from the special loop, supporting the notion that the special loop represents a site where ADAR1 is stored, possibly for later use during development.
Sallacz
Nina B
NB
Department of Chromosome Biology, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria.
Jantsch
Michael F
MF
D000243:Adenosine Deaminase / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D002875:Chromosomes / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D003609:Dactinomycin / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D017403:In Situ Hybridization; D017404:In Situ Hybridization, Fluorescence; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009865:Oocytes / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D017393:RNA Editing; D016601:RNA-Binding Proteins; D012260:Ribonucleases / Q000378:metabolism; D014158:Transcription, Genetic; D014982:Xenopus laevis / Q000378:metabolism*
D017931:DNA Primers; D016601:RNA-Binding Proteins; D016222:Fibroblast Growth Factor 2; D003609:Dactinomycin; D012313:RNA; D012260:Ribonucleases; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase; D001973:Bromodeoxyuridine