Nucleic Acids Research, Vol 27, Issue 17 3455-3465, Copyright © 1999 by Oxford University Press
BG Luukkonen and B Seraphin
A combination of point mutations disrupting both stem 1 and stem 2 of U5
snRNA (U5AI) was found to confer a thermosensitive phenotype in vivo. In a
strain expressing U5AI, pre-mRNA splicing was blocked before the first step
through an inability of the mutant U5 snRNA to efficiently associate with
the U4/U6 di-snRNP. Formation of early splicing complexes was not affected
in extracts prepared from U5 snRNA mutant cells, while the capacity of
these extracts to splice a pre-mRNA in vitro was greatly diminished. In
addition, significant levels of a translation product derived from intron
containing pre-mRNAs could be detected in vivo. The SSD1/SRK1 gene was
identified as a multi-copy suppressor of the U5AI snRNA mutant. Single copy
expression of SSD1/SRK1 was sufficient to suppress the thermosensitive
phenotype, and high copy expression partially suppressed the splicing and
U4/U6.U5 tri- snRNP assembly pheno-types. SSD1/SRK1 also suppressed
thermosensitive mutations in the Prp18p and U1-70K proteins, while
inhibiting growth of the cold sensitive U1-4U snRNA mutant at 30 degrees C.
Thus we have identified SSD1/SRK1 as a general suppressor of splicing
mutants.
ARTICLES
A conditional U5 snRNA mutation affecting pre-mRNA splicing and nuclear pre-mRNA retention identifies SSD1/SRK1 as a general splicing mutant suppressor
European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
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