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
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.