Human U3 small nucleolar RNA genes are localized to the nucleoplasm

Abstract

U3 RNA, an abundant, conserved, capped, small RNA localized to the nucleolar compartment of eukaryotic cells, is implicated in the processing of pre-ribosomal RNA. The genes for U1 and U2 snRNAs ( s mall n uclear RNAs) are clustered and present in the nucleoplasmic DNA; however, the localization of U3 snRNA genes is not known. DNAs, isolated from HeLa cell nuclei and nucleoli, were hybridized with labeled probes corresponding to 1) the 5′-flanking and 2) the coding regions of the human U3 snRNA gene. The intensity of signals obtained with both the probes were 5–6 fold greater in nuclear DNA, compared to nucleolar DNA. With ribosomal gene probe, the nucleolar DNA had sixfold more intense signal than nuclear DNA. These results indicate that genes for U3 snRNA are in the nucleoplasm. Therefore, U3 snRNA, like 5S ribosomal RNA, appears to be synthesized in the nucleoplasm and transported to the nucleolus.