B. Lubben et al., ISOLATION AND CHARACTERIZATION OF THE SMALL NUCLEOLAR RIBONUCLEOPROTEIN PARTICLE SNR30 FROM SACCHAROMYCES-CEREVISIAE, The Journal of biological chemistry, 270(19), 1995, pp. 11549-11554
The nucleolus of the yeast Saccharomyces cerevisiae contains the small
nucleolar RNA snR30 (snoRNA), that is found associated with at least
two proteins, NOP1 and GAR1. Ah three of these molecules are essential
for the cell's viability and have been implicated in pre-rRNA maturat
ion. NOP1 and GAR1 are believed to be general rRNA-processing factors
or, alternatively, integral protein components of the small nucleolar
ribonucleoprotein particle snR30 (snoRNP). In this paper, we describe
procedures for the biochemical isolation of snR30 RNP, and we identify
seven snR30 RNP proteins of molecular masses of 10, 23, 25, 38, 46, 4
8, and 65 kDa, including the previously reported GAR1 protein. Additio
nal proteins, including NOP1, may also be components of snR30 RNP but
are lost during our stringent isolation procedure. The 10, 23-, and 25
-kDa (GAR1) and 65-kDa proteins remain tightly associated with the snR
30 RNA even after isopycnic sedimentation in cesium sulfate gradients.
Electron microscopy of Mono and-purified snR30 RNPs show a slightly e
longated two-domain structure approximately 20 nm long and 14 nm wide.