The RNA-editing enzyme ADAR1 is localized to the nascent ribonucleoprotein matrix on Xenopus lampbrush chromosomes but specifically associates with an atypical loop.
Double-stranded RNA adenosine deaminase (ADAR1, dsRAD, DRADA) converts adenosines to inosines in double-stranded RNAs. Few candidate substrates for ADAR1 editing are known at this point and it is not known how substrate recognition is achieved. In some cases editing sites are defined by basepaired regions formed between intronic and exonic sequences, suggesting that the enzyme might function cotranscriptionally. We have isolated two variants of Xenopus laevis ADAR1 for which no editing substrates are currently known. We demonstrate that both variants of the enzyme are associated with transcriptionally active chromosome loops suggesting that the enzyme acts cotranscriptionally. The widespread distribution of the protein along the entire chromosome indicates that ADAR1 associates with the RNP matrix in a substrate-independent manner. Inhibition of splicing, another cotranscriptional process, does not affect the chromosomal localization of ADAR1. Furthermore, we can show that the enzyme is dramatically enriched on a special RNA-containing loop that seems transcriptionally silent. Detailed analysis of this loop suggests that it might represent a site of ADAR1 storage or a site where active RNA editing is taking place. Finally, mutational analysis of ADAR1 demonstrates that a putative Z-DNA binding domain present in ADAR1 is not required for chromosomal targeting of the protein.
Eckmann
C R
CR
Department of Cytology and Genetics, Institute of Botany, University of Vienna, A-1030 Vienna, Austria.
Jantsch
M F
MF
D000243:Adenosine Deaminase / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D001665:Binding Sites; D002875:Chromosomes / Q000201:enzymology* / Q000648:ultrastructure; D014644:Genetic Variation; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D011499:Protein Processing, Post-Translational; D017393:RNA Editing; D012326:RNA Splicing; D016601:RNA-Binding Proteins; D011817:Rabbits; D012261:Ribonucleoproteins / Q000378:metabolism*; D014982:Xenopus laevis
D000906:Antibodies; D009838:Oligodeoxyribonucleotides; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase