Cloning and characterization of P110, a novel small nucleolar U3 ribonucleoprotein, expressed in early development.

ABSTRACT

We report the cloning of a BrdU-sensitive transcript of 4.1 kb from an immortalized quail heart cell line containing an open reading frame of 940 amino acids (110 kDa, pI approximately 5.18). The mRNA encoding P110 appears in the heart and neural tube by 36 h of avian development, at a time when these organs are rapidly developing. Analysis of the DNA-deduced protein sequence revealed a bipartite nuclear localization signal, and a highly charged domain rich in both acidic and basic residues. Immunofluorescent staining with polyclonal antibodies raised against a P110 peptide localized the protein to the nucleolus of avian and mammalian cells. Although database search showed significant homology with an uncharacterized cDNA from human brain and several human and mouse Expressed Sequence Tags, there was no close homology to known nucleolar proteins. Immunoprecipitation of P110 from cell sonicates revealed it contained U3 small nucleolar RNA, but no significant amounts of other box C/D small nucleolar RNAs. These data suggest that P110 is one of the U3 small nucleolar ribonucleoproteins that are involved in rRNA processing.