Identification of antisense RNA transcripts from a human DNA topoisomerase I pseudogene

Abstract

Eukaryotic topoisomerase I (TOP1), a DNA unwinding enzyme, plays an essential role in several cellular functions; however, regulation of TOP1 activity remains unknown. In an effort to identify potential regulators of TOP1 activity, the transcriptional activity of a TOP1 pseudogene in chromosome 1 was studied. By using primers unique to the TOP1 pseudogene, strand-specific polymerase chain reaction analysis of HeLa RNA amplified products from at least two transcripts oriented in the antisense direction of TOP1 mRNA. In one case, polymerase chain reaction and Northern blot analysis found a 0.7-kilobase antisense transcript. Upon estimation of 5' and 3' boundaries, a 497 base stretch of homology with the TOP1 mRNA was found. While the function of these TOP1 antisense transcripts remains unknown, recent studies of naturally occurring antisense RNA have demonstrated several potential regulatory roles. The production of antisense transcripts from a TOP1 pseudogene was the first example of a naturally occurring antisense RNA transcript produced from a pseudogene.