Purification of the yeast U4/U6 center dot U5 small nuclear ribonucleoprotein particle and identification of its proteins

Citation
Sw. Stevens et J. Abelson, Purification of the yeast U4/U6 center dot U5 small nuclear ribonucleoprotein particle and identification of its proteins, P NAS US, 96(13), 1999, pp. 7226-7231
Citations number
63
Categorie Soggetti
Multidisciplinary
Journal title
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
ISSN journal
00278424 → ACNP
Volume
96
Issue
13
Year of publication
1999
Pages
7226 - 7231
Database
ISI
SICI code
0027-8424(19990622)96:13<7226:POTYUC>2.0.ZU;2-L
Abstract
The yeast U4/U6 . U5 pre-mRNA splicing small nuclear ribonucleoprotein (snR YP) is a 25S small nuclear ribonucleoprotein particle similar in size, comp osition, and morphology to its counterpart in human cells, The yeast U4/U6 . U5 snRYP complex has been purified to near homogeneity by. affinity chrom atography and preparative glycerol gradient sedimentation. We show that the re are at least 24 proteins stably associated with this particle and perfor med mass spectrometry microsequencing to determine their identities. In add ition to the seven canonical core Sm proteins, there are a set of U6 snRSP specific Sm proteins, eight previously described U4/U6 . U5 snRNP proteins, and four novel proteins. Two of the novel proteins have likely RNA binding properties, one has been implicated in the cell cycle and one has no ident ifiable sequence homologues or functional motifs, The purification of the l ow abundance U4/U6 . U5 snRYP from yeast and the powerful sequencing method ologies using small amounts of protein make possible the rapid identificati on of novel and previously unidentified components of large, low-abundance macromolecular machines from any genetically manipulable organism.